Antigen binding proteins capable of binding thymic stromal lymphopoietin

ABSTRACT

The present disclosure provides compositions and methods relating to antigen binding proteins which bind to human thymic stromal lymphopoietin (TSLP), including antibodies. In particular embodiments, the disclosure provides fully human, humanized and chimeric anti-TSLP antibodies and derivatives of such antibodies. The disclosure further provides nucleic acids encoding such antibodies and antibody fragments and derivatives, and methods of making and using such antibodies including methods of treating and preventing TSLP-related inflammatory and fibrotic disorders.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §119 of U.S.Provisional Application Ser. No. 61/091,676, filed Aug. 25, 2008 andU.S. Provisional Application Ser. No. 60/971,178 filed Sep. 10, 2007,which are hereby incorporated by reference.

FIELD OF THE INVENTION

The field of this invention relates to compositions of antigen bindingproteins including antibodies capable of binding human thymic stromallymphopoietin, as well as related methods.

BACKGROUND OF THE INVENTION

The prevalence of allergic diseases such as asthma, allergic rhinitis,atopic dermatitis, and food allergies appears to be increasing in recentyears, particularly in developed countries, affecting an increasingpercentage of the population (Kay, N Engl. J. Med. 344:30-37 (2001)).Thymic stromal lymphopoietin (TSLP) is an epithelial cell derivedcytokine produced in response to pro-inflammatory stimuli. TSLP has beendiscovered to promote allergic inflammatory responses primarily throughits activity on dendritic and mast cells (Soumelis et al., Nat Immun3(7): 673-680 (2002), Allakhverdi et al., J. Exp. Med. 204(2):253-258(2007)). Human TSLP expression has been reported to be increased inasthmatic airways correlating to disease severity (Ying et al., J.Immunol. 174: 8183-8190 (2005)). In addition, TSLP protein levels aredetectable in the concentrated bronchoalveoloar lavage (BAL) fluid ofasthma patients, and other patients suffering from allergic disorders.Also, increased levels of TSLP protein and mRNA are found in thelesional skin of atopic dermatitis (AD) patients. Therefore, TSLPantagonists would be useful in treating inflammatory disorders.

In addition, TSLP has also been found to promote fibrosis, as reportedin U.S. application Ser. No. 11/344,379. Fibrotic disease results duringthe tissue repair process if the fibrosis phase continues unchecked,leading to extensive tissue remodeling and the formation of permanentscar tissue (Wynn, Nature Rev. Immunol. 4, 583 (2004)). It has beenestimated that up to 45% of deaths in the United States can beattributed to fibroproliferative diseases, which can affect many tissuesand organ systems (Wynn, supra, at 595 (2004)).

Currently, anti-inflammatory treatments are used to treat fibroticdisorders, since fibrosis is common to many persistent inflammatorydiseases such as idopathic pulmonary fibrosis, progressive kidneydisease, and liver cirrhosis. However, the mechanisms involved inregulation of fibrosis appear to be distinctive from those ofinflammation, and anti-inflammatory therapies are not always effectivein reducing or preventing fibrosis (Wynn, supra). Therefore, a needremains for developing treatments to reduce and prevent fibrosis.

Therefore, antagonists to TSLP would be expected to be useful fortreating these inflammatory and fibrotic disorders. The presentdisclosure provides such treatments and methods of treating.

SUMMARY OF THE INVENTION

In one aspect, the present disclosure provides an isolated antigenbinding protein comprising a. a light chain CDR3 sequence selected fromi. a light chain CDR3 sequence that differs by no more than a total oftwo amino acid additions, substitutions, and/or deletions from a CDR3sequence selected from the group consisting of the light chain CDR3sequences of A1 to A27; ii. QQAX₈SFPLT (SEQ ID NO: 251); and b. a heavychain CDR3 sequence selected from i. a heavy chain CDR3 sequence thatdiffers by no more than a total of three amino acid additions,substitutions, and/or deletions from a CDR3 sequence selected from thegroup consisting of the heavy chain CDR3 sequences of A1 to A27; ii.GGGIX₁₂VADYYX₁₃YGMDV (SEQ ID NO: 255); iii. DX₂₁GX₂₂SGWPLFX₂₃Y (SEQ IDNO: 259); wherein X₈ is an N residue or a D residue; X₁₂ is a P residueor an A residue; X₁₃ is a Y residue or an F residue; X₂₁ is a G residueor an R residue; X₂₂ is an S residue or a T residue; X₂₃ is an A residueor a D residue, and wherein said antigen binding protein specificallybinds to TSLP.

In another aspect, the isolated antigen binding protein of the presentdisclosure further comprises at least one of the following: a. a lightchain CDR1 sequence selected from i. a light chain CDR1 sequence thatdiffers by no more than three amino acids additions, substitutions,and/or deletions from a light chain CDR1 sequence of A1-A27; ii.RSSQSLX₁YSDGX₂TYLN (SEQ ID NO: 246);

iii. RASQX₄X₅SSWLA (SEQ ID NO: 249); b. a light chain CDR2 sequenceselected from i. a light chain CDR2 sequence that differs by no morethan two amino acid additions, substitutions, and/or deletions from aCDR2 sequence of A1-A27; ii. KVSX₃ (residues 1-4 of SEQ ID NO: 247);iii. X₆X₇SSLQS (SEQ ID NO: 250); or iv. QDX₉KRPS (SEQ ID NO: 252); andc. a heavy chain CDR1 sequence selected from i. a heavy chain CDR1sequence that differs by no more than two amino acid additions,substitutions, and/or deletions from a CDR1 sequence of A1-A27; ii.X₁₀YGMH (SEQ ID NO: 253); and iii. X₁₅X₁₆YMX₁₇ (SEQ ID NO: 257); and d.a heavy chain CDR2 sequence selected from i. a heavy chain CDR2 sequencethat differs by no more than three amino acid additions, substitutions,and/or deletions from a CDR2 sequence of A1-A27; ii. VIWX₁₁DGSNKYYADSVKG(SEQ ID NO: 254); iii. VISYDGSX₁₄KYYADSVKG (SEQ ID NO: 256); and iv.WINPNSGGTNX₁₈X₁₉X₂₀KFQG (SEQ ID NO: 258); wherein X₁ is a V residue oran I residue; X₂ is an N residue or a D residue; X₃ is a Y residue or anN residue; X₄ is a G residue or a S residue; X₅ is a L residue or an Iresidue; X₆ is an N residue or a T residue; X₇ is a T residue or an Aresidue; X₉ is a K residue or an N residue; X₁₀ is an S residue or an Nresidue; X₁₁ is a Y residue or an F residue; X₁₄ is a Y residue or a Nresidue; X₁₅ is a D residue or G residue; X₁₆ is a Y residue or a Dresidue; X₁₇ is a Y residue or an H residue; X₁₈ is a Y residue or an Hresidue; X₁₉ is a V residue or an A residue; X₂₀ is a Q residue or an Rresidue, and wherein said antigen binding protein specifically binds toTSLP.

In another aspect of the present disclosure, the isolated antigenbinding protein of claim 1 comprises either: a. a light chain variabledomain comprising: i. a light chain CDR1 sequence selected from A1-A27;ii a light chain CDR2 sequence selected from A1-A27; iii. a light chainCDR3 sequence selected from A1-A27; or b. a heavy chain variable domaincomprising i. a heavy chain CDR1 sequence selected from A1-A27; ii. aheavy chain CDR2 sequence selected from A1-A27, and iii. a heavy chainCDR3 sequence selected from A1-A27; or c. the light chain variabledomain of (a) and the heavy chain variable domain of (b).

In a further aspect, the isolated antigen binding protein compriseseither a. a light chain variable domain sequence selected from i. aminoacids having a sequence at least 80% identical to a light chain variabledomain sequence selected from L1-L27; ii. a sequence of amino acidsencoded by a polynucleotide sequence that is at least 80% identical to apolynucleotide sequence encoding the light chain variable domainsequence of L1-L27; iii. a sequence of amino acids encoded by apolynucleotide sequence that hybridizes under moderately stringentconditions to the complement of a polynucleotide consisting of a lightchain variable domain sequence of L1-L27; b. a heavy chain variabledomain sequence selected from i. a sequence of amino acids that is atleast 80% identical to a heavy chain variable domain sequence of H1-H27;ii. a sequence of amino acids encoded by a polynucleotide sequence thatis at least 80% identical to a polynucleotide sequence encoding theheavy chain variable domain sequence of H1-H27; iii. a sequence of aminoacids encoded by a polynucleotide sequence that hybridizes undermoderately stringent conditions to the complement of a polynucleotideconsisting of a heavy chain variable domain sequence of H1-H27; or c.the light chain variable domain of (a) and the heavy chain variabledomain of (b), wherein said antigen binding protein specifically bindsto TSLP.

In a further aspect, an isolated antigen binding protein of the presentdisclosure comprises either: a. a light chain variable domain sequenceselected from: L1-L27; b. a heavy chain variable domain sequenceselected from H1-H27; or, c. the light chain variable domain of (a) andthe heavy chain variable domain of (b), wherein the antigen bindingprotein specifically binds to TSLP.

In a further aspect, the isolated binding protein comprises a lightchain variable domain sequence and a heavy chain variable domainsequence selected from L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8,L9H9, L10H10, L11H11, L12H12, L13.1H13, L13.2H13, L14.1H14, L14.2H14,L15.1H15, L15.2H15, L16.1H16, L16.2H16, L17H17, L18.1H18, L18.2H18,L19.1H19, L19.2H19, L20.1H20, L20.2H20, L21H21, L22H22, L23H23, L24H24,L25H25, L26H26, and L27H27.

In a further aspect, the isolated antigen binding protein comprises abinding protein that binds to TSLP with substantially the same Kd as areference antibody selected from A2, A3, A4, and A5. In another aspect,the isolated antigen binding protein comprises a binding protein thatinhibits TSLP activity according to the primary cell OPG assay with thesame IC₅₀ as a reference antibody selected from A2, A3, A4 or A5.

In a still further aspect, the isolated antigen binding proteincross-competes for binding of TSLP with a reference antibody. In anotheraspect, the isolated antigen binding protein binds the same epitope as areference antibody, e.g., A2, A4, A5, A6, A7, A₁₀, A21, A23, or A26.

In one aspect, the isolated antigen binding protein is selected from ahuman antibody, a humanized antibody, chimeric antibody, a monoclonalantibody, a polyclonal antibody, a recombinant antibody, anantigen-binding antibody fragment, a single chain antibody, a diabody, atriabody, a tetrabody, a Fab fragment, an F(fa′)x fragment, a domainantibody, an IgD antibody, an IgE antibody, and IgM antibody, and IgG1antibody, and IgG2 antibody, and IgG3 antibody, and IgG4 antibody, andIgG4 antibody having at least one mutation in the hinge region thatalleviates a tendency to for intra H-chain disulfide bonds. In oneaspect, the isolated antigen binding protein is a human antibody.

Also provided is an isolated nucleic acid molecule comprising apolynucleotide sequence encoding the light chain variable domain, theheavy chain variable domain, or both, of the antigen binding agent ofthe present disclosure. In one embodiment, the polynucleotide comprisesa light chain variable sequence L1-L27, and/or a heavy chain variablesequence H1-H27, or both.

Also provided are vectors comprising the polynucleotides of the presentdisclosure. In one embodiment the vector is an expression vector. Alsoprovided is a host cell comprising the vector. Also provided is ahybridoma capable of producing the antigen binding protein of thepresent invention. Also provided is a method of making the antigenbinding protein comprising culturing the host cell under conditions thatallow it to express the antigen binding protein.

Also provided is a pharmaceutical composition comprising the antigenbinding proteins of the present invention. In one embodiment thepharmaceutical composition comprises a human antibody. Also provided isa method of treating a TSLP-related inflammatory condition in a subjectin need of such treatment comprising administering a therapeuticallyeffective amount of the composition to the subject. In one embodiment,the inflammatory condition is allergic asthma, allergic rhinosinusitis,allergic conjunctivitis, or atopic dermatitis. Also provided is a methodof treating a TSLP-related fibrotic disorder in a subject in need ofsuch treatment comprising administering a therapeutically effectiveamount of the composition to the subject. In one embodiment, thefibrotic disorder is scleroderma, interstitial lung disease, idiopathicpulmonary fibrosis, fibrosis arising from chronic hepatitis B or C,radiation-induced fibrosis, and fibrosis arising from wound healing.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A-FIG. 1F. The figure provides the amino acid sequence of thelight chain CDR1, CDR2, and CDR3 regions of A1-A27. Further provided isan exemplary nucleotide sequence encoding each CDR.

FIG. 2A-FIG. 2F. The figure provides the amino acid sequence of theheavy chain CDR1, CDR2, and CDR3 regions of A1-A27. Further provided isan exemplary nucleotide sequence encoding each CDR.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to antigen binding agents, includingantigen binding proteins, that specifically bind to the cytokine humanthymic stromal lymphopoietin (TSLP), including antigen binding proteinsthat inhibit TSLP binding and signaling such as antagonistic TSLPantibodies, antibody fragments, and antibody derivatives. The antigenbinding agents are useful for inhibiting or blocking binding of TSLP toits receptor, and for treating inflammatory diseases, fibrotic diseases,and other related conditions.

The present invention further provides compositions, kits, and methodsrelating to antigen binding proteins that bind to TSLP. Also providedare nucleic acid molecules, and derivatives and fragments thereof,comprising a sequence of polynucleotides that encode all or a portion ofa polypeptide that binds to TSLP, such as a nucleic acid encoding all orpart of an anti-TSLP antibody, antibody fragment, or antibodyderivative. The present invention further provides vectors and plasmidscomprising such nucleic acids, and cells or cell lines comprising suchnucleic acids and/or vectors and plasmids. The provided methods include,for example, methods of making, identifying, or isolating antigenbinding proteins that bind to human TSLP such as anti-TSLP antibodies,methods of determining whether an antigen binding protein binds to TSLP,methods of making compositions, such as pharmaceutical compositions,comprising an antigen binding protein that binds to TSLP, and methodsfor administering an antigen binding protein that binds to TSLP in asubject, for example, methods for treating a condition mediated by TSLP,and for modulating a biological activity associated with TSLP signallingin vivo or in vitro.

TSLP

Thymic stromal lymphopoietin (TSLP) refers to a four α-helical bundletype I cytokine which is a member of the IL-2 family but most closelyrelated to IL-7. Cytokines are low molecular weight regulatory proteinssecreted in response to certain stimuli, which act on receptors on themembrane of target cells. Cytokines regulate a variety of cellularresponses. Cytokines are generally described in references such asCytokines, A. Mire-Sluis and R. Thorne, ed., Academic Press, New York,(1998).

TSLP was originally cloned from a murine thymic stromal cell line (Simset al J. Exp. Med. 192 (5), 671-680 (2000)), and found to support earlyB and T cell development. Human TSLP was later cloned and found to havea 43 percent identity in amino acid sequence to the murine homolog(Quentmeier et al. Leukemia 15, 1286-1292 (2001), and U.S. Pat. No.6,555,520, which is herein incorporated by reference). Thepolynucleotide and amino acid sequence of human TSLP are presented inSEQ ID NO: 1 and 2 respectively. TSLP was found to bind with lowaffinity to a receptor chain from the hematopoietin receptor familycalled TSLP receptor (TSLPR), which is described in U.S. patentapplication Ser. No. 09/895,945 (publication No: 2002/0068323) (SEQ IDNO: 3 and 4). The polynucleotide sequence encoding human TSLPR ispresented as SEQ ID NO: 3 of the present application, and the amino acidsequence is presented as SEQ ID NO: 4 of the present applicationrespectively. The soluble domain of the TSLPR is approximately aminoacids 25 through 231 of SEQ ID NO: 4. TSLP binds with high affinity to aheterodimeric complex of TSLPR and the interleukin 7 receptor alphaIL-7Rα (Park et al., J. Exp. Med. 192:5 (2000), U.S. patent applicationSer. No. 09/895,945, publication number U.S. 2002/0068323). The sequenceof IL-7 receptor α is shown in FIG. 2 of U.S. Pat. No. 5,264,416, whichis herein incorporated by reference. The sequence of the soluble domainof the IL-7 receptor α is amino acid 1 to 219 of FIG. 2 in U.S. Pat. No.5,264,416.

As used herein the term “TSLP polypeptides” refers to various forms ofTSLP useful as immunogens. These include TSLP expressed in modifiedform, in which a furin cleavage site has been removed throughmodification of the amino acid sequence, as described in PCT patentapplication publication WO 03/032898. Modified TSLP retains activity butthe full length sequence is more easily expressed in mammalian cellssuch as CHO cells. Examples of TSLP polypeptides include SEQ ID NO: 2,SEQ ID NO: 373, and SEQ ID NO: 375.

In addition, cynomolgus TSLP has been identified and is shown in Example1 below and is set forth in SEQ ID NO: 380, for example.

TSLP is produced in human epithelial cells including skin, bronchial,tracheal, and airway epithelial cells, keratinocytes, stromal and mastcells, smooth muscle cells, and lung and dermal fibroblasts, asdetermined by quantitative mRNA analysis (Soumelis et al, NatureImmunol. 3 (7) 673-680 (2002)). Both murine and human TSLP are involvedin promoting allergic inflammation.

TABLE 1 Database(s) (or Protein Patent Accession Name Species SynonymsApplication) No. SLP Homo sapiens Thymic stromal GenBank/ AAK67940/lymphopoietin protein SEQ ID NO: 2 of U.S. Pat. No. 6,555,520 ModifiedHomo sapiens Thymic stromal SEQ ID NOS: 10, TSLP lymphopoietin 12, 14,16, 17, 18 of WO 03/032898 TSLP Mus musculus Thymic stroma derivedGenBank AAF81677 lymphopoietin; Thymic stromal derived lymphopoietinTSLPR Homo sapiens Cytokine receptor-like 2 SEQ ID NO: 5 of (CRL2);IL-XR; Thymic US 2002/0068323 stromal lymphopoietin protein receptorTSLPR Mus Cytokine receptor-like GenBank, Q8CII9 factor 2; Type Icytokine SWISSPROT receptor delta 1; Cytokine receptor-like molecule 2(CRLM-2); Thymic stromal lymphopoietin protein receptor IL-7R Homosapiens Interleukin-7 receptor GenBank/ NM_002185 U.S. Pat. No.:5,264,416

TSLP Activity

TSLP activities include the proliferation of BAF cells expressing humanTSLPR (BAF/HTR), as described in PCT patent application publication WO03/032898. The BAF/HTR bioassay utilizes a murine pro B lymphocyte cellline, which has been transfected with the human TSLP receptor. TheBAF/HTR cells are dependent upon huTSLP for growth, and proliferate inresponse to active huTSLP added in test samples. Following an incubationperiod, cell proliferation is measured by the addition of Alamar Bluedye I or tritiated thymidine. Proliferation may also be measured using acommercially available kit such as the CYQUANT cell proliferation assaykit (Invitrogen).

Additional assays for huTSLP activity include, for example, an assaymeasuring induction of T cell growth from human bone marrow by TSLP asdescribed in U.S. Pat. No. 6,555,520. Another TSLP activity is theability to activate STAT5 as described in the reference to Levin et al.,J. Immunol. 162:677-683 (1999) and PCT patent application WO 03/032898.

Additional assays include TSLP induced CCL17/TARC production fromprimary human monocytes and dendritic cells as described in USapplication publication no. 2006/0039910 (Ser. No. 11/205,909).

Cell based assays useful for measuring TSLP activity are described inthe examples below. These include the BAF cell proliferation assaydescribed above, as well as the primary cell assay described belowmeasuring TSLP induced osteoprotegerin (OPG) production from primaryhuman dendritic cells, as well cynomolgus peripheral blood mononuclearcell assay, also described below.

TSLP activities further include in vivo activities. These can bemeasured in mouse models, for example, such as those described in Zhouet al., Nat Immunol 6(10), 1047-1053 (2005), and Yoo et al., J Exp Med.202 (4), 541-549 (2005). For example, an anti-murine TSLP antibody wasshown to decrease BALF cellularity and BALF levels of IL-5 and Il-13 inan Ova-asthma model (Zhou et al).

Definitions

Polynucleotide and polypeptide sequences are indicated using standardone- or three-letter abbreviations. Unless otherwise indicated,polypeptide sequences have their amino termini at the left and theircarboxy termini at the right, and single-stranded nucleic acidsequences, and the top strand of double-stranded nucleic acid sequences,have their 5′ termini at the left and their 3′ termini at the right. Aparticular polypeptide or polynucleotide sequence also can be describedby explaining how it differs from a reference sequence.

Polynucleotide and polypeptide sequences of particular light and heavychain variable domains., L1 (“light chain variable domain 1”), H1(“heavy chain variable domain 1”), etc. Antibodies comprising a lightchain and heavy chain are indicated by combining the name of the lightchain and the name of the heavy chain variable domains. For example,“L4H7,” indicates an antibody comprising the light chain variable domainof L4 and the heavy chain variable domain of H7.

Unless otherwise defined herein, scientific and technical terms used inconnection with the present invention shall have the meanings that arecommonly understood by those of ordinary skill in the art. Further,unless otherwise required by context, singular terms shall includepluralities and plural terms shall include the singular. Generally,nomenclatures used in connection with, and techniques of, cell andtissue culture, molecular biology, immunology, microbiology, geneticsand protein and nucleic acid chemistry and hybridization describedherein are those well known and commonly used in the art. The methodsand techniques of the present invention are generally performedaccording to conventional methods well known in the art and as describedin various general and more specific references that are cited anddiscussed throughout the present specification unless otherwiseindicated. See, e.g., Sambrook et al. Molecular Cloning: A LaboratoryManual, 2d ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor,N.Y. (1989) and Ausubel et al., Current Protocols in Molecular Biology,Greene Publishing Associates (1992), and Harlow and Lane Antibodies: ALaboratory Manual Cold Spring Harbor Laboratory Press, Cold SpringHarbor, N.Y. (1990), which are incorporated herein by reference.Enzymatic reactions and purification techniques are performed accordingto manufacturer's specifications, as commonly accomplished in the art oras described herein. The terminology used in connection with, and thelaboratory procedures and techniques of, analytical chemistry, syntheticorganic chemistry, and medicinal and pharmaceutical chemistry describedherein are those well known and commonly used in the art. Standardtechniques can be used for chemical syntheses, chemical analyses,pharmaceutical preparation, formulation, and delivery, and treatment ofpatients.

The following terms, unless otherwise indicated, shall be understood tohave the following meanings: The term “isolated molecule” (where themolecule is, for example, a polypeptide, a polynucleotide, or anantibody) is a molecule that by virtue of its origin or source ofderivation (1) is not associated with naturally associated componentsthat accompany it in its native state, (2) is substantially free ofother molecules from the same species (3) is expressed by a cell from adifferent species, or (4) does not occur in nature. Thus, a moleculethat is chemically synthesized, or expressed in a cellular systemdifferent from the cell from which it naturally originates, will be“isolated” from its naturally associated components. A molecule also maybe rendered substantially free of naturally associated components byisolation, using purification techniques well known in the art. Moleculepurity or homogeneity may be assayed by a number of means well known inthe art. For example, the purity of a polypeptide sample may be assayedusing polyacrylamide gel electrophoresis and staining of the gel tovisualize the polypeptide using techniques well known in the art. Forcertain purposes, higher resolution may be provided by using HPLC orother means well known in the art for purification.

The terms “TSLP inhibitor” and “TSLP antagonist” are usedinterchangeably. Each is a molecule that detectably inhibits TSLPsignalling. The inhibition caused by a TSLP inhibitor need not becomplete so long as it is detectable using an assay. For example, thecell-based assay described in Example 4 below, demonstrates an assayuseful for determining TSLP signaling inhibition.

The terms “peptide” “polypeptide” and “protein” each refers to amolecule comprising two or more amino acid residues joined to each otherby peptide bonds. These terms encompass, e.g., native and artificialproteins, protein fragments and polypeptide analogs (such as muteins,variants, and fusion proteins) of a protein sequence as well aspost-translationally, or otherwise covalently or non-covalently,modified proteins. A peptide, polypeptide, or protein may be monomericor polymeric.

The term “polypeptide fragment” as used herein refers to a polypeptidethat has an amino-terminal and/or carboxy-terminal deletion as comparedto a corresponding full-length protein. Fragments can be, for example,at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 50, 70, 80, 90, 100,150 or 200 amino acids in length. Fragments can also be, for example, atmost 1,000, 750, 500, 250, 200, 175, 150, 125, 100, 90, 80, 70, 60, 50,40, 30, 20, 15, 14, 13, 12, 11, or 10 amino acids in length. A fragmentcan further comprise, at either or both of its ends, one or moreadditional amino acids, for example, a sequence of amino acids from adifferent naturally-occurring protein (e.g., an Fc or leucine zipperdomain) or an artificial amino acid sequence (e.g., an artificial linkersequence).

Polypeptides of the invention include polypeptides that have beenmodified in any way and for any reason, for example, to: (1) reducesusceptibility to proteolysis, (2) reduce susceptibility to oxidation,(3) alter binding affinity for forming protein complexes, (4) alterbinding affinities, and (4) confer or modify other physicochemical orfunctional properties. Analogs include muteins of a polypeptide. Forexample, single or multiple amino acid substitutions (e.g., conservativeamino acid substitutions) may be made in the naturally occurringsequence (e.g., in the portion of the polypeptide outside the domain(s)forming intermolecular contacts). A “conservative amino acidsubstitution” is one that does not substantially change the structuralcharacteristics of the parent sequence (e.g., a replacement amino acidshould not tend to break a helix that occurs in the parent sequence, ordisrupt other types of secondary structure that characterize the parentsequence or are necessary for its functionality). Examples ofart-recognized polypeptide secondary and tertiary structures aredescribed in Proteins, Structures and Molecular Principles (Creighton,Ed., W. H. Freeman and Company, New York (1984)); Introduction toProtein Structure (C. Branden and J. Tooze, eds., Garland Publishing,New York, N.Y. (1991)); and Thornton et al. Nature 354:105 (1991), whichare each incorporated herein by reference.

A “variant” of a polypeptide comprises an amino acid sequence whereinone or more amino acid residues are inserted into, deleted from and/orsubstituted into the amino acid sequence relative to another polypeptidesequence. Variants of the invention include fusion proteins. Variants ofantibodies described herein also include those that result fromprocessing. Such variants include those having one, two, three, four,five, six, seven, eight, nine ten or more additional amino acids at theN-terminus of a light or heavy chain, e.g., as a result of inefficientsignal sequence cleavage. Such variants also include those missing oneor more amino acids from the N- or C-termini of a light or heavy chain.

A “derivative” of a polypeptide is a polypeptide (e.g., an antibody)that has been chemically modified, e.g., via conjugation to anotherchemical moiety such as, for example, polyethylene glycol, albumin(e.g., human serum albumin), phosphorylation, and glycosylation. Unlessotherwise indicated, the term “antibody” includes, in addition toantibodies comprising two full-length heavy chains and two full-lengthlight chains, derivatives, variants, fragments, and muteins thereof,examples of which are described below.

An “antigen binding protein” according to the present disclosure is aprotein capable of binding to an antigen and, optionally, a scaffold orframework portion that allows the antigen binding portion to adopt aconformation that promotes binding of the antigen binding protein to theantigen. In one embodiment an antigen binding protein of the presentinvention comprises at least one CDR. Examples of antigen bindingproteins include antibodies, antibody fragments (e.g., an antigenbinding portion of an antibody), antibody derivatives, and antibodyanalogs. The antigen binding protein can comprise, for example, analternative protein scaffold or artificial scaffold with grafted CDRs orCDR derivatives. Such scaffolds include, but are not limited to,antibody-derived scaffolds comprising mutations introduced to, forexample, stabilize the three-dimensional structure of the antigenbinding protein as well as wholly synthetic scaffolds comprising, forexample, a biocompatible polymer. See, for example, Korndorfer et al.,2003, Proteins: Structure, Function, and Bioinformatics, Volume 53,Issue 1:121-129; Roque et al., 2004, Biotechnol. Prog. 20:639-654. Inaddition, peptide antibody mimetics (“PAMs”) can be used, as well asscaffolds based on antibody mimetics utilizing fibronection componentsas a scaffold.

An antigen binding protein can have, for example, the structure of anaturally occurring immunoglobulin. An “immunoglobulin” is a tetramericmolecule. In a naturally occurring immunoglobulin, each tetramer iscomposed of two identical pairs of polypeptide chains, each pair havingone “light” (about 25 kDa) and one “heavy” chain (about 50-70 kDa). Theamino-terminal portion of each chain includes a variable region of about100 to 110 or more amino acids primarily responsible for antigenrecognition. The carboxy-terminal portion of each chain defines aconstant region primarily responsible for effector function. Human lightchains are classified as kappa and lambda light chains. Heavy chains areclassified as mu, delta, gamma, alpha, or epsilon, and define theantibody's isotype as IgM, IgD, IgG, IgA, and IgE, respectively. Withinlight and heavy chains, the variable and constant regions are joined bya “J” region of about 12 or more amino acids, with the heavy chain alsoincluding a “D” region of about 10 more amino acids. See generally,Fundamental Immunology Ch. 7 (Paul, W., ed., 2^(nd) ed. Raven Press,N.Y. (1989)) (incorporated by reference in its entirety for allpurposes). The variable regions of each light/heavy chain pair form theantibody binding site such that an intact immunoglobulin has two bindingsites.

Naturally occurring immunoglobulin chains exhibit the same generalstructure of relatively conserved framework regions (FR) joined by threehypervariable regions, also called complementarity determining regionsor CDRs. From N-terminus to C-terminus, both light and heavy chainscomprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Theassignment of amino acids to each domain is in accordance with thedefinitions of Kabat et al. in Sequences of Proteins of ImmunologicalInterest, 5^(th) Ed., US Dept. of Health and Human Services, PHS, NIH,NIH Publication no. 91-3242, 1991. Intact antibodies include polyclonal,monoclonal, chimeric, humanized or fully human having full length heavyand light chains.

An “antibody” refers to an intact immunoglobulin or to an antigenbinding portion thereof that competes with the intact antibody forspecific binding, unless otherwise specified. Antigen binding portionsmay be produced by recombinant DNA techniques or by enzymatic orchemical cleavage of intact antibodies. Antigen binding portions includeFab, Fab′, F(ab′)₂, Fd, Fv, and domain antibodies (dAbs), andcomplementarity determining region (CDR) fragments, single-chainantibodies (scFv), diabodies, triabodies, tetrabodies, and polypeptidesthat contain at least a portion of an immunoglobulin that is sufficientto confer specific antigen binding to the polypeptide.

A Fab fragment is a monovalent fragment having the V_(L), V_(H), C_(L)and C_(H)1 domains; a F(ab′)₂ fragment is a bivalent fragment having twoFab fragments linked by a disulfide bridge at the hinge region; a Fdfragment has the V_(H) and C_(H)1 domains; an Fv fragment has the V_(L)and V_(H) domains of a single arm of an antibody; and a dAb fragment hasa V_(H) domain, a V_(L) domain, or an antigen-binding fragment of aV_(H) or V_(L) domain (U.S. Pat. Nos. 6,846,634, 6,696,245, US App. Pub.No. 05/0202512, 04/0202995, 04/0038291, 04/0009507, 03/0039958, Ward etal., Nature 341:544-546, 1989).

A single-chain antibody (scFv) is an antibody in which a V_(L) and aV_(H) region are joined via a linker (e.g., a synthetic sequence ofamino acid residues) to form a continuous protein chain wherein thelinker is long enough to allow the protein chain to fold back on itselfand form a monovalent antigen binding site (see, e.g., Bird et al.,1988, Science 242:423-26 and Huston et al., 1988, Proc. Natl. Acad. Sci.USA 85:5879-83). Diabodies are bivalent antibodies comprising twopolypeptide chains, wherein each polypeptide chain comprises V_(H) andV_(L) domains joined by a linker that is too short to allow for pairingbetween two domains on the same chain, thus allowing each domain to pairwith a complementary domain on another polypeptide chain (see, e.g.,Holliger et al., 1993, Proc. Natl. Acad. Sci. USA 90:6444-48, and Poljaket al., 1994, Structure 2:1121-23). If the two polypeptide chains of adiabody are identical, then a diabody resulting from their pairing willhave two identical antigen binding sites. Polypeptide chains havingdifferent sequences can be used to make a diabody with two differentantigen binding sites. Similarly, tribodies and tetrabodies areantibodies comprising three and four polypeptide chains, respectively,and forming three and four antigen binding sites, respectively, whichcan be the same or different.

Complementarity determining regions (CDRs) and framework regions (FR) ofa given antibody may be identified using the system described by Kabatet al. in Sequences of Proteins of Immunological Interest, 5^(th) Ed.,US Dept. of Health and Human Services, PHS, NIH, NIH Publication no.91-3242, 1991. One or more CDRs may be incorporated into a moleculeeither covalently or noncovalently to make it an antigen bindingprotein. An antigen binding protein may incorporate the CDR(s) as partof a larger polypeptide chain, may covalently link the CDR(s) to anotherpolypeptide chain, or may incorporate the CDR(s) noncovalently. The CDRspermit the antigen binding protein to specifically bind to a particularantigen of interest.

An antigen binding protein may have one or more binding sites. If thereis more than one binding site, the binding sites may be identical to oneanother or may be different. For example, a naturally occurring humanimmunoglobulin typically has two identical binding sites, while a“bispecific” or “bifunctional” antibody has two different binding sites.

The term “human antibody” includes all antibodies that have one or morevariable and constant regions derived from human immunoglobulinsequences. In one embodiment, all of the variable and constant domainsare derived from human immunoglobulin sequences (a fully humanantibody). These antibodies may be prepared in a variety of ways,examples of which are described below, including through theimmunization with an antigen of interest of a mouse that is geneticallymodified to express antibodies derived from human heavy and/or lightchain-encoding genes.

A humanized antibody has a sequence that differs from the sequence of anantibody derived from a non-human species by one or more amino acidsubstitutions, deletions, and/or additions, such that the humanizedantibody is less likely to induce an immune response, and/or induces aless severe immune response, as compared to the non-human speciesantibody, when it is administered to a human subject. In one embodiment,certain amino acids in the framework and constant domains of the heavyand/or light chains of the non-human species antibody are mutated toproduce the humanized antibody. In another embodiment, the constantdomain(s) from a human antibody are fused to the variable domain(s) of anon-human species. In another embodiment, one or more amino acidresidues in one or more CDR sequences of a non-human antibody arechanged to reduce the likely immunogenicity of the non-human antibodywhen it is administered to a human subject, wherein the changed aminoacid residues either are not critical for immunospecific binding of theantibody to its antigen, or the changes to the amino acid sequence thatare made are conservative changes, such that the binding of thehumanized antibody to the antigen is not significantly worse than thebinding of the non-human antibody to the antigen. Examples of how tomake humanized antibodies may be found in U.S. Pat. Nos. 6,054,297,5,886,152 and 5,877,293.

The term “chimeric antibody” refers to an antibody that contains one ormore regions from one antibody and one or more regions from one or moreother antibodies. In one embodiment, one or more of the CDRs are derivedfrom a human anti-TSLP antibody. In another embodiment, all of the CDRsare derived from a human anti-TSLP antibody. In another embodiment, theCDRs from more than one human anti-TSLP antibodies are mixed and matchedin a chimeric antibody. For instance, a chimeric antibody may comprise aCDR1 from the light chain of a first human anti-TSLP antibody, a CDR2and a CDR3 from the light chain of a second human anti-TSLP antibody,and the CDRs from the heavy chain from a third anti-TSLP antibody.Further, the framework regions may be derived from one of the sameanti-TSLP antibodies, from one or more different antibodies, such as ahuman antibody, or from a humanized antibody. In one example of achimeric antibody, a portion of the heavy and/or light chain isidentical with, homologous to, or derived from an antibody from aparticular species or belonging to a particular antibody class orsubclass, while the remainder of the chain(s) is/are identical with,homologous to, or derived from an antibody (-ies) from another speciesor belonging to another antibody class or subclass. Also included arefragments of such antibodies that exhibit the desired biologicalactivity (i.e., the ability to specifically bind the human TSLPreceptor).

Fragments or analogs of antibodies can be readily prepared by those ofordinary skill in the art following the teachings of this specificationand using techniques well-known in the art. Preferred amino- andcarboxy-termini of fragments or analogs occur near boundaries offunctional domains. Structural and functional domains can be identifiedby comparison of the nucleotide and/or amino acid sequence data topublic or proprietary sequence databases. Computerized comparisonmethods can be used to identify sequence motifs or predicted proteinconformation domains that occur in other proteins of known structureand/or function. Methods to identify protein sequences that fold into aknown three-dimensional structure are known. See, e.g., Bowie et al.,1991, Science 253:164.

A “CDR grafted antibody” is an antibody comprising one or more CDRsderived from an antibody of a particular species or isotype and theframework of another antibody of the same or different species orisotype.

A “multi-specific antibody” is an antibody that recognizes more than oneepitope on one or more antigens. A subclass of this type of antibody isa “bi-specific antibody” which recognizes two distinct epitopes on thesame or different antigens.

An antigen binding protein including an antibody “specifically binds” toan antigen, such as TSLP if it binds to the antigen with a high bindingaffinity as determined by a Kd (or corresponding Kb, as defined below)value of 10⁻⁷ M or less.

An “antigen binding domain,” “antigen binding region,” or “antigenbinding site” is a portion of an antigen binding protein that containsamino acid residues (or other moieties) that interact with an antigenand contribute to the antigen binding protein's specificity and affinityfor the antigen. For an antibody that specifically binds to its antigen,this will include at least part of at least one of its CDR domains.

The “percent identity” of two polynucleotide or two polypeptidesequences is determined by comparing the sequences using the GAPcomputer program (a part of the GCG Wisconsin Package, version 10.3(Accelrys, San Diego, Calif.)) using its default parameters.

The terms “polynucleotide,” “oligonucleotide” and “nucleic acid” areused interchangeably throughout and include DNA molecules (e.g., cDNA orgenomic DNA), RNA molecules (e.g., mRNA), analogs of the DNA or RNAgenerated using nucleotide analogs (e.g., peptide nucleic acids andnon-naturally occurring nucleotide analogs), and hybrids thereof. Thenucleic acid molecule can be single-stranded or double-stranded. In oneembodiment, the nucleic acid molecules of the invention comprise acontiguous open reading frame encoding an antibody, or a fragment,derivative, mutein, or variant thereof, of the invention.

Two single-stranded polynucleotides are “the complement” of each otherif their sequences can be aligned in an anti-parallel orientation suchthat every nucleotide in one polynucleotide is opposite itscomplementary nucleotide in the other polynucleotide, without theintroduction of gaps, and without unpaired nucleotides at the 5′ or the3′ end of either sequence. A polynucleotide is “complementary” toanother polynucleotide if the two polynucleotides can hybridize to oneanother under moderately stringent conditions. Thus, a polynucleotidecan be complementary to another polynucleotide without being itscomplement.

A “vector” is a nucleic acid that can be used to introduce anothernucleic acid linked to it into a cell. One type of vector is a“plasmid,” which refers to a linear or circular double stranded DNAmolecule into which additional nucleic acid segments can be ligated.Another type of vector is a viral vector (e.g., replication defectiveretroviruses, adenoviruses and adeno-associated viruses), whereinadditional DNA segments can be introduced into the viral genome. Certainvectors are capable of autonomous replication in a host cell into whichthey are introduced (e.g., bacterial vectors comprising a bacterialorigin of replication and episomal mammalian vectors). Other vectors(e.g., non-episomal mammalian vectors) are integrated into the genome ofa host cell upon introduction into the host cell, and thereby arereplicated along with the host genome. An “expression vector” is a typeof vector that can direct the expression of a chosen polynucleotide.

A nucleotide sequence is “operably linked” to a regulatory sequence ifthe regulatory sequence affects the expression (e.g., the level, timing,or location of expression) of the nucleotide sequence. A “regulatorysequence” is a nucleic acid that affects the expression (e.g., thelevel, timing, or location of expression) of a nucleic acid to which itis operably linked. The regulatory sequence can, for example, exert itseffects directly on the regulated nucleic acid, or through the action ofone or more other molecules (e.g., polypeptides that bind to theregulatory sequence and/or the nucleic acid). Examples of regulatorysequences include promoters, enhancers and other expression controlelements (e.g., polyadenylation signals). Further examples of regulatorysequences are described in, for example, Goeddel, 1990, Gene ExpressionTechnology: Methods in Enzymology 185, Academic Press, San Diego, Calif.and Baron et al., 1995, Nucleic Acids Res. 23:3605-06.

A “host cell” is a cell that can be used to express a nucleic acid,e.g., a nucleic acid of the invention. A host cell can be a prokaryote,for example, E. coli, or it can be a eukaryote, for example, asingle-celled eukaryote (e.g., a yeast or other fungus), a plant cell(e.g., a tobacco or tomato plant cell), an animal cell (e.g., a humancell, a monkey cell, a hamster cell, a rat cell, a mouse cell, or aninsect cell) or a hybridoma. Exemplary host cells include Chinesehamster ovary (CHO) cell lines or their derivatives including CHO strainDXB-11, which is deficient in DHFR (see Urlaub et al., 1980, Proc. Natl.Acad. Sci. USA 77:4216-20), CHO cell lines which grow in serum-freemedia (see Rasmussen et al., 1998, Cytotechnology 28:31), CS-9 cells, aderivative of DXB-11 CHO cells, and AM-1/D cells (described in U.S. Pat.No. 6,210,924). Other CHO cells lines include CHO-K1 (ATCC# CCL-61), EM9(ATCC# CRL-1861), and UV20 (ATCC# CRL-1862). Examples of other hostcells include COS-7 line of monkey kidney cells (ATCC CRL 1651) (seeGluzman et al., 1981, Cell 23:175), L cells, C127 cells, 3T3 cells (ATCCCCL 163), HeLa cells, BHK (ATCC CRL 10) cell lines, the CV1/EBNA cellline derived from the African green monkey kidney cell line CV1 (ATCCCCL 70) (see McMahan et al., 1991, EMBO J. 10:2821), human embryonickidney cells such as 293, 293 EBNA or MSR 293, human epidermal A431cells, human Colo205 cells, other transformed primate cell lines, normaldiploid cells, cell strains derived from in vitro culture of primarytissue, primary explants, HL-60, U937, HaK or Jurkat cells. Typically, ahost cell is a cultured cell that can be transformed or transfected witha polypeptide-encoding nucleic acid, which can then be expressed in thehost cell. The phrase “recombinant host cell” can be used to denote ahost cell that has been transformed or transfected with a nucleic acidto be expressed. A host cell also can be a cell that comprises thenucleic acid but does not express it at a desired level unless aregulatory sequence is introduced into the host cell such that itbecomes operably linked with the nucleic acid. It is understood that theterm host cell refers not only to the particular subject cell but to theprogeny or potential progeny of such a cell. Because certainmodifications may occur in succeeding generations due to, e.g., mutationor environmental influence, such progeny may not, in fact, be identicalto the parent cell, but are still included within the scope of the termas used herein.

Antigen Binding Proteins

In one aspect, the present disclosure provides antigen binding proteinssuch as antibodies, antibody fragments, antibody derivatives, antibodymuteins, and antibody variants that bind to human TSLP. Antigen bindingproteins in accordance with the present disclosure includes antigenbinding proteins that bind to human TSLP, and thereby reduce TSLPactivity. For example, antigen binding proteins may interfere with thebinding of TSLP to its receptor, and thus reduce TSLP activity.

In one embodiment, the present invention provides an antigen bindingprotein that comprises one or more CDR sequences that differ from a CDRsequence shown in FIG. 1A-1F or FIG. 2A-2F by no more than 5, 4, 3, 2,1, or 0 amino acid residues.

In another embodiment, at least one of the antigen binding protein CDR3sequence is a sequence from FIG. 1A-1F or FIG. 2A-2F. In anotherembodiment, the antigen binding protein's light chain CDR3 sequence is alight chain sequence from A1 through A27, and the antigen bindingprotein heavy chain CDR3 sequence is a heavy chain CDR3 sequence from A1through A27.

In another embodiment, the antigen binding protein further comprises 1,2, 3, 4, or 5 CDR sequences that each independently differs by 5, 4, 3,2, 1, or 0 single amino acid additions, substitutions, and/or deletionsfrom a CDR sequence of A1-A27. The light chain CDR's of exemplaryantigen binding proteins A1-A27 and the heavy chain CDR's of exemplarybinding proteins A1-A27 are shown in FIG. 1A-1F and FIG. 2A-2F,respectively. Also shown are polynucleotide sequences which encode theamino acid sequences of the CDRs. In addition, consensus sequences ofthe CDR sequences are provided below.

CDR CONSENSUS SEQUENCES VARIABLE LIGHT CHAIN CDRs Group 1a LC CDR1Consensus             X₁        X₂ A16.1 R S S Q S L V Y S D G N T Y L NA18.1             V         N A13.1             V         D A19.1            V         D A20.1             V         D A14.1            V         N A15.1             I         N R S S Q S L X₁ Y SD G X₂ T Y L N (SEQ ID NO: 246) X₁ is a V (valine) residue or an I(isoleucine) residue, X₂ is an N (asparagine) residue or a D (aspartic)acid residue; LC CDR2 Consensus       X₃ A16.1 K V S Y W D S A18.1      Y A13.1       N A19.1       N A20.1       N A14.1       N A15.1      N KVSX₃WDS (SEQ ID NO: 247) X₃ is a Y (tyrosine) residue or an N(asparagine) residue; LC CDR3 consensus A16.1 M Q G T H W P P A A18.1A13.1 A19.1 A20.1 A14.1 A15.1 MQGTHQPPA (SEQ ID NO: 248) Group 1b LCCDR1 consensus         X₄ X₅ A13.2 R A S Q G  L S S W L A A14.2        G  L A19.2         G  L A20.2         G  L A16.2         S  LA18.2         S  L A15.2         G  I RASQX₄X₅SSWLA (SEQ ID NO: 249) X₄is a G (glycine) residue or an S (serine) residue; X₅ is a L (leucine)residue or an I (isoleucine) residue; LC CDR2 consensus X₆ X₇ A13.2 N  T S S L Q S A14.2 N  T A19.2 N  T A20.2 N  T A16.2 N  A A18.2 N  A A15.2T  T X₆X₇SSLQS (SEQ ID NO: 250) X6 is an N (asparagine) residue or a T(threonine) residue; X7 is a T(theonine) residue or an A (alanine)residue; LC CDR3 consensus       X₈ A13.2 Q Q A N S F P L T A14.2      N A19.2       N A20.2       N A16.2       N A18.2       N A15.2      D QQAX₈SFPLT (SEQ ID NO: 251) X₈ is a N (asparagine) residue or aD (aspartic acid) residue; Group 2 LC CDR1 consensus A6 S G D K L G D KY A C A8 SGDKLGDKYAC (SEQ ID NO: 15) LC CDR2 consensus     X A6 Q D K KR P 5 A8     N QDX₉KRPS (SEQ ID NO: 252) X₉ is a K (lysine) residue oran N (asparagine) residue; LC CDR3 consensus A6 Q A W D S S T V V A8QAWDSSTVV (SEQ ID NO: 107) Group 3 LC CDR1 consensus A3 T G S S S N I GA G F D V H A4 TGSSSNIGAGFDVH (SEQ ID NO: 10) LC CDR2 consensus A3 D N NN R P S A4 DNNNRPS (SEQ ID NO: 57) LC CDR3 consensus A3 Q S Y D S N L SG S I V V A4 QSYDSNLSGSIVV (SEQ ID NO: 102) VARIABLE HEAVY CHAIN CDRSGroup 1 HC CDR1 consensus X₁₀ A13 S  Y G M H A14 S A19 S A20 S A16 N A18N A15 N X₁₀YGMH (SEQ ID NO: 253) X₁₀ is a S (serine) or an N(asparagine) residue; HC CDR2 consensus       X₁₁ A13 V I W Y  D G S N KY Y A D S V K G A14       Y A19       Y A20       Y A16       Y A18      Y A15       F VIWX₁₁DGSNKYYADSVKG (SEQ ID NO: 254) X₁₁ is a Y(tyrosine) residue or a F (phenyl- alanine) residue. HC CDR3 consensus        X₁₂           X₁₃ A13 G G G I P  V A D Y Y Y Y G M D V A14        P            Y A19         P            Y A20        P            Y A16         A            Y A18        A            Y A15         A            F GGGIX₁₂VADYYX₁₃YGMDV(SEQ ID NO: 255) X₁₂ is a P (proline) residue or an A (alanine) residue;X₁₃ is a Y (tyrosine) residue or a F (phenyl- alanine) residue. Group 2HC CDR1 consensus A6 S Y G I H A8 SYGIH (SEQ ID NO: 147) HC CDR2consensus               X₁₄ A6 V I S Y D G S Y   K Y Y A D S V K G A8              N VISYDGSX₁₄KYYADSVKG (SEQ ID NO: 256) X₁₄ is a Y(tyrosine) or an N (asparagine) residue. HC CDR3 consensus A6 G D S W ND R L N Y Y F Y D M D V A8 GDSWNDRLNYYFYDMDV (SEQ ID NO: 214) Group 3 HCCDR1 consensus X₁₅ X₁₆     X₁₇ A3 D  Y  Y M Y A4 G  D      H X₁₅X₁₆YMX₁₇(SEQ ID NO: 257) X₁₅ is a D (aspartic acid) or G (glycine) residue; X₁₆is a Y (tyrosine) or D (aspartic acid) residue; X₁₇ is a Y (tyrosine) oran H (histidine) residue. HC CDR2 consensus                     X₁₈ X₁₉X₂₀ A3 W I N P N S G G T N Y  V  Q K F Q G A4                    H  A  R WINPNSGGTNX₁₈X₁₉X₂₀KFQG (SEQ ID NO: 258) X₁₈is a Y (tyrosine) or H (histidine) residue; X₁₉ is a V (valine) or A(alanine) residue; X₂₀ is a Q (glutamine) or R (arginine) residue. HCCDR3 consensus   X₂₁  X₂₂           X₂₃ A3 D G  G S   G W P L F A  Y A4  R    T             D (SEQ ID NO: 259) X₂₁ is a G (glycine) or R(arginine) residue; X₂₂ is a S (serine) or T (threonine) residue; X₂₃ isan A (alanine) or D (aspartic acid) residue.

Table 2 below provides nucleic acid (DNA) sequences encoding thevariable heavy domains (H#) and variable light domains (L#), and theamino acid sequences of the variable heavy and variable light domainsfor exemplary TSLP antigen binding proteins A1-A27, respectively. CDRs1, 2 & 3 for each variable domain are sequential from the beginning tothe end of each sequence. Framework (Fr) regions are underlined.Frameworks 1, 2, 3 & 4 for each variable domain are sequential from thebeginning to the end of each sequence (e.g., the first underlinedportion of the sequence is Fr1, the second is Fr2, the third is Fr3 &the last is Fr4 in each sequence).

TABLE 2 H1 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:260) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTATATTACTGTGCGAGTCTAGTGGGAGCTACCAACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCG TCTCCTCA H1Protein QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVIWYDGSNKY (SEQID NO: 261) YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCASLVGATNYYGMDVWGQGTTVTVSS L1 DNA TCTTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATC(SEQ ID NO: 262)ACATGCCAAGGAGACAGCCTCAGAAGCTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTCTGGTAAAAACTACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTACTACTGTAACTCCCGGGACAGAAGTGGTAACCATCTGGTGTTTTCGGCGGAGGGACCAAGCTGACCGTCCTA L1 ProteinSSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVISGKNYRPSGIPDRFSG (SEQ IDNO: 263) SSSGNTASLTITGAQAEDEADYYCNSRDRSGNHLVFGGGTKLTVL H2 DNAGAAGTGCAGCTGGTGGAGTCTGGGGGAGTCGTGGTACAGCCTGGGGGGTCCCTGAGACT (SEQ ID NO:264) CTCCTGTGCAGCCTCTGGATTCACCTTTGATGATTTTACCATGCACTGGGTCCGTCAAGCTCCGGGGAAGGGTCTGGAGTGGGTCTCTCTTATTAGTTGGGATGGTGGTAGCACATACTATGCAGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAACAGCAAAAACTCCCTGTATATGCAAATGAACAGTCTGAGAACTGAGGACAGCGCCTTGTATTACTGTGCAAGAGGTCCTTACTACTACTTCTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCT CA H2Protein EVQLVESGGVVVQPGGSLRLSCAASGFTFDDFTMHWVRQAPGKGLEWVSLISWDGGSTYY(SEQ ID NO: 265)ADSVKGRFTISRDNSKNSLYMQMNSLRTEDSALYYCARGPYYYFYGMDVWGQGTTVTVSS L2 DNATCTTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATC (SEQ ID NO:266) ACATGCCAAGGAGACAGCCTCAGAACCTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTATACTTGTCATCTCTGATAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGGGACAGCAGTGATAACCATGTAGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L2 ProteinSSELTQDPAVSVALGQTVRITCQGDSLRTYYASWYQQKPGQAPILVISDKNNRPSGIPDRFSG (SEQ IDNO: 267) SSSGNTASLTITGAQAEDEADYYCNSRDSSDNHLVVFGGGTKLTVL H3 DNACAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGGCTGGGGCCTCAGTGAAGGT (SEQ ID NO:268) CTCCTGCAAGGCTTCTGGATACACCTTCACCGACTACTATATGTACTGGGTGCGACAGGCCCCTGGACAAGGGCCTGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACTATGTACAGAAGTTTCAGGGCAGGGTCACCATGACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGATGAGATCCGACGACACGGCCGTGTATTACTGTGCGAGAGATGGGGGTAGCAGTGGCTGGCCCCTCTTTGCCTACTGGGGCCTGGGAACCCTGGTCACCGT CTCCTCA H3Protein QVQLVQSGAEVKKPGASVKVSCKASGYTFTDYYMYWVRQAPGQGPEWMGWINPNSGGTN (SEQID NO: 269) YVQKFQGRVTMTRDTSISTAYMELSRMRSDDTAVYYCARDGGSSGWPLFAYWGLGTLVTVSS L3 DNA CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCAT(SEQ ID NO: 270)CTCCTGCACTGGGAGCAGCTCGAACATCGGGGCAGGTTTTGATGTACACTGGTACCAGCAGCTTCCAGGAACAGCCCCCAAACTCCTCATCTATGATAACAACAATCGGCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAGCAACCTGAGTGGTTCGATTGTGGTTTTTCGGCGGAGGGACCAAGCTGACCGTCCTA L3 ProteinQSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQQLPGTAPKLLIYDNNNRPSGVPDR (SEQ IDNO: 271) FSGSKSGTSASLAITGLQAEDEADYYCQSYDSNLSGSIVVFGGGTKLTVL H4 DNACAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGT (SEQ ID NO:272) CTCCTGCAAGGCTTCTGGATACATCTTCACCGGCGACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTGGAGTGGATGGGATGGATCAACCCTAACAGTGGTGGCACAAACCATGCACGGAAGTTTCAGGGCAGGGTCACCATGACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGATCTGACGACACGGCCGTGTATTACTGTGTGAGAGATAGGGGTACCAGTGGCTGGCCACTCTTTTGACTATTGGGGCCAGGGAACACTGGTCACCGT CTCCTCA H4Protein QVQLVQSGAEVKKPGASVKVSCKASGYIFTGDYMHWVRQAPGQGLEWMGWINPNSGGTN (SEQID NO: 273) HARKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCVRDRGTSGWPLFDYWGQGTLVTVSS L4 DNA CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCAT(SEQ ID NO: 274)CTCCTGCACTGGGAGCAGCTCCAACATCGGGGCAGGTTTTGATGTGCACTGGTACCAGCTGCTTCCAGGAACAGCCCCCAAACTCCTCATCTTTGATAACAACAATCGCCCCTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGGCTCCAGGCTGAGGATGAGGCTGATTATTACTGCCAGTCCTATGACAGCAACCTGAGTGGTTCGATTGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L4 ProteinQSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGFDVHWYQLLPGTAPKLLIFDNNNRPSGVPDR (SEQ IDNO: 275) FSGSKSGTSASLAITGLQAEDEADYYCQSYDSNLSGSIVVFGGGTKLTVL H5 DNACAGATGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:360) CTCCTGTGCAGCGTCTGGATTCACCTTCAGAACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAACACTATGCAGACTCCGTGAAGGGCCGATTCACCATCACCAGAGACAATTCCAAGAACACTCTGAATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGCCCCTCAGTGGGAGCTAGTTCATGAAGCTTTTGATATCTGGGGCCAAGGGACAATGGTCAC CGTCTCTTCAH5 Protein QMQLVESGGGVVQPGRSLRLSCAASGFTFRTYGMHWVRQAPGKGLEWVAVIWYDGSNKH(SEQ ID NO: 361)YADSVKGRFTITRDNSKNTLNLQMNSLRAEDTAVYYCARAPQWELVHEAFDIWGQGTMVT VSS L5 DNATCCTATGTGCTGACTCAGCCACCCTCGGTGTCAGTGGCCCCAGGACAGACGGCCAGGATT (SEQ ID NO:362) ACCTGTGGGGGAAACAACCTTGGAAGTAAAAGTGTGCACTGGTACCAGCAGAAGCCAGGCCAGGCCCCTGTGCTGGTCGTCTATGATGATAGCGACCGGCCCTCATGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAGCAGGGGCGAAGCCGGGGATGAGGCCGACTATTACTGTCAGGTGTGGGATAGTAGTAGTGATCATGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L5 ProteinSYVLTQPPSVSVAPGQTARITCGGNNLGSKSVHWYQQKPGQAPVLVVYDDSDRPSWIPERFS (SEQ IDNO: 363) GSNSGNTATLTISRGEAGDEADYYCQVWDSSSDHVVFGGGTKLTVL H6 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:276) CTCCTGTGCAGCCTCTGGATTCATTTTCAGTAGCTATGGCATTCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTTATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGACTCCTGGAACGACAGATTAAACTACTACTTCTACGATATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H6 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFIFSSYGIHWVRQAPGKGLEWVAVISYDGSYKYYA (SEQ IDNO: 277) DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGDSWNDRLNYYFYDMDVWGQGTTVTVSS L6 DNATCCTATGAGCTGACTCAGGCACCCTCAGTGTCCGTGTCCCCAGGACAGACAGCCAGCATC (SEQ ID NO:278) ACCTGCTCTGGAGATAAATTGGGGGATAAATATGCTTGCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTGCTGGTCATCTATCAAGATAAGAAGCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACAGCCACTCTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGCAGCACTGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L6 ProteinSYELTQAPSVSVSPGQTASITCSGDKLGDKYACWYQQKPGQSPVLVIYQDKKRPSGIPERFSG (SEQ IDNO: 279) SNSGNTATLTISGTQAMDEADYYCQAWDSSTVVFGGGTKLTVL H7 DNACAGGTGCAGCTGCAGGAGTCGGGCCCAGGACTGGTGAAGCCTTCACAGACCCTGTCCCT (SEQ ID NO:280) CACCTGGACTGTCTCTGGTGGCTCCATCAGCAGTGGTGGTTACTACTGGAGCTGGATCCGCCAGCACCCAGGGAAGGGCCTGGAGTGGATTGGGTTCATCCATTACAGTGGGACCACCTACTACAACCCGTCCCTCAAGAGTCGACTTACCCTATCAGTAGACACGTCTAAGAGCCAGTTCTCCCTGAAGCTGAACTCTGTGACTGCCGCGGACACGGCCGTGTATTACTGTGCGAGAGAAGTTGGCAGCTCGTCGGGTAACTGGTTCGACCCCTGGGGCCAGGGAACCCTGGTCACC GTCTCCTCA H7Protein QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGGYYWSWIRQHPGKGLEWIGFIHYSGTTYYNP(SEQ ID NO: 281)SLKSRLTLSVDTSKSQFSLKLNSVTAADTAVYYCAREVGSSSGNWFDPWGQGTLVTVSS L7 DNATCCTATGAGCTGACTCAGCCACCCTCAGTGTCCGTGTCCCCAGGACAGACAGCCAGCATC (SEQ ID NO:282) ACCTGCTCTGGAGATAAATTGGGGGATAAATATGCTTGCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTGGTGGTCATCTATCAAGATAACAAGCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACAGCCACTTTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGCACCACTGCGATATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L7 ProteinSYELTQPPSVSVSPGQTASITCSGDKLGDKYACWYQQKPGQSPVVVIYQDNKRPSGIPERFSG (SEQ IDNO: 283) SNSGNTATLTISGTQAMDEADYYCQAWDSTTAIFGGGTKLTVL H8 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:284) CTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGCTATGGCATTCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATCATATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGACTCCTGGAACGACAGATTAAACTACTACTTCTACGATATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H8 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGIHWVRQAPGKGLEWVAVISYDGSNKYYA (SEQ IDNO: 285) DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGDSWNDRLNYYFYDMDVWGQGTTVTVSS L8 DNATCCTATGAGCTGACTCAGCCACCCTCAGTGTCCGTGTCCCCAGGACAGACAGCCAGCATC (SEQ ID NO:286) ACCTGCTCTGGAGATAAATTGGGGGATAAATATGCTTGCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTACTGGTCATCTATCAAGATAACAAGCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACAGCCACTTTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGCAGCACTGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L8 ProteinSYELTQPPSVSVSPGQTASITCSGDKLGDKYACWYQQKPGQSPVLVIYQDNKRPSGIPERFSG (SEQ IDNO: 287) SNSGNTATLTISGTQAMDEADYYCQAWDSSTVVFGGGTKLTVL H9 DNACAGGTGCAGTTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:288) CTCCTGTGCAGCGTCTGGATATACCTTCAATAGCTATGGCATGCACTGGGTGCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATACATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACATTTCCAAGAACACTCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAGGTCCGGGCGTATAGCAGTGGCTGGTACGCCGCCTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA H9 ProteinQVQLVESGGGVVQPGRSLRLSCAASGYTFNSYGMHWVRQAPGKGLEWVAVIWYDGSNTY (SEQ ID NO:289) YADSVKGRFTISRDISKNTLYLQMNSLRAEDTAVYYCAREVRAYSSGWYAAFDYWGQGTL VTVSSL9 DNA TCTTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATC (SEQID NO: 290) ACATGCCAAGGAGACAGCCTCAGAATCTTTTATGCAAACTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTAGTTGTCTTCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGCGGCTCAGGCGGAAGATGAGGCTGACTATTATTGTAACTCCCGGGACAGCAGTGGTAACCATGTGGTATTTCGGCGGAGGGACCACGCTGACCGTCCTA L9 ProteinSSELTQDPAVSVALGQTVRITCQGDSLRIFYANWYQQKPGQAPVVVFYGKNNRPSGIPDRFS (SEQ IDNO: 291) GSSSGNTASLTITAAQAEDEADYYCNSRDSSGNHVVFGGGTTLTVL H10 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:292) CTCCTGTGCAACGTCTGGATTCACCTTCAGTAGTTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAGTAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTGTATTACTGTGCGAGAGTAAGAAGTGGGAGCTACTACGAACAGTATTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCGCCGTCTCCTCA H10 ProteinQVQLVESGGGVVQPGRSLRLSCATSGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSSKYY (SEQ ID NO:293) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVRSGSYYEQYYYGMDVWGQGTT VAVSSL10 DNA GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACC(SEQ ID NO: 294)ATCACTTGCCGGGCAAATCAGTACATTAGCACCTATTTAAATTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGGTCCTGATTTATGCTGGATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATTTGAGACAGATTTCACTCTCACCATCAGCAGTCTGCAACCTGAAGATTTTGCAACTTACTACTGTCAGCAGAGCTACACTACCCCGATCACCTTTCGGCCAAGGGACACGACTGGAGATTAAA L10 ProteinDIQMTQSPSSLSASVGDRVTITCRANQYISTYLNWYQQKPGKAPKVLIYAASSLQSGVPSRFS (SEQ IDNO: 295) GSGFETDFTLTLSSLQPEDFATYYCQQSYTTPITFGQGTRLEIK H11 DNAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACT (SEQ ID NO:296) CTCCTGTGCAGCCTCTGGATTCACCTTCAGTAGTTATAGCATGAACTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTTTCATACATTAGTGGTCGTACTAGTAGCGTATACTACGCAGACTCTGTGAAGGGCCGATTCACCATCTCCAGAGACAATGCCAAGAACTCACTGTATCTGCACATGAACAGCCTGAGAGACGAGGACACGGCTGTGTATTACTGTGCGAGAAGTGGGATCTACTACGACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGT CTCCTCA H11Protein EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYSMNWVRQAPGKGLEWVSYISGRTSSVYYA(SEQ ID NO: 297)DSVKGRFTISRDNAKNSLYLHMNSLRDEDTAVYYCARSGIYYDYYGMDVWGQGTTVTVSS L11 DNAGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCCCC (SEQ ID NO:298) ATCAACTGCAAGTCCAGCCAGAGTGTTTTAAACAGCTCCAACAATAAGAACTACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGACATCCACCCGGGAAGGCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAAGATGTGGCAGTTTATTACTGTCAGCAGTATTTTACTACTCCGTGGACGTTTCGGCCAAGGGAGCAAGGTGGAGATCAAA L11 ProteinDIVMTQSPDSLAVSLGERAPINCKSSQSVLNSSNNKNYLAWYQQKPGQPPKLLIYWTSTREG (SEQ IDNO: 299) GVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQQYFTTPWTFGQGTKVEIK H12 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:300) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGCAGCCACTGCTATAGATTACTACTACTCCTACGGTATGGACGTCTGGGGCCTAGGGACCACGGTCACCGTCTCCTCA H12 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:301) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGAATAIDYYYSYGMDVWGLGTT VTVSSL12 DNA GACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTGGGAGACAGAGTCACC(SEQ ID NO: 302)ATCACTTGTCGGGCGAGTCAGGGTATTAGTAGCTGGTIAGCCTGGTATCAGCGGAAACCAGGAAAAGCCCCTAAGTTCCTGATCTATACTGCATCCAGTTTGCAAAGTGGGGTCCCATCACGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTCTGCAACTTACTATTGTCAACAGGCTGACAGTTTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA L12 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQRKPGKAPKFLIYTASSLQSGVPSRFS (SEQ IDNO: 303) GSGSGTDFTLTISSLQPEDSATYYCQQADSFPLTFGGGTKVEIK H13 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:304) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATACCAGTAGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCGTCA H13 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:305) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIPVADYYYYGMDVWGQGTT VTVSSL13.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 306)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTCTACAGTGATGGAGACACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTAACTGGGACTCTGGGGTCCCATACAGATTCAGCGGCAGTGGGTCAGGCACTGATTTCACACTGCAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGATTTACTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L13.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGDTYLNWFQQRPGQSPRRLIYKVSNWDSG (SEQ IDNO: 307) VPYRFSGSGSGTDFTLQISRVEAEDVGIYYCMQGTHWPPAFGQGTRLEIK L13.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGAGAGAGTCACC (SEQ ID NO:308) ATCACTTGTCGGGCGAGTCAGGGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCCTGATGTATAACACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAGTTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA L13.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGLSSWLAWYQQKPGKAPKLLMYNTSSLQSGVPSRF (SEQ IDNO: 309) SGSGSGTDFSLTISSLQPEDFASYYCQQANSFPLTFGGGTKVEIK H14 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:304) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATACCAGTAGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H14 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:305) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIPVADYYYYGMDVWGQGTT VTVSSL14.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 310)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTCTACAGTGATGGAAACACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTAACTGGGACTCTGGGGTCCCAGACAGATTCAGCGGCATTGGGTCAGGCACTGACTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGGTTTACTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L14.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGNTYLNWFQQRPGQSPRRLIYKVSNWDSG (SEQ IDNO: 311) VPDRFSGIGSGTDFTLKISRVEAEDVGVYYCMQGTHWPPAFGQGTRLEIK L14.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:312) ATCACTTGTCGGGCGAGTCAGGGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCCTGATGTATAACACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAGTTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA L14.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGLSSWLAWYQQKPGKAPKLLMYNTSSLQSGVPSRF (SEQ IDNO: 309) SGSGSGTDFSLTISSLQPEDFASYYCQQANSFPLTFGGGTKVEIK H15 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAAGTCCCTGAGACT (SEQ ID NO:313) CTCCTGTGCAGCGTCTGGATTCCCCTTCAGTAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTGGAATGGGTGGCAGTTATATGGTTTGATGGAAGTAATAAATACTATGCGGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATCCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATAGCAGTGGCTGACTACTACTTCTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H15 ProteinQVQLVESGGGVVQPGKSLRLSCAASGFPFSNYGMHWVRQAPGKGLEWVAVIWFDGSNKYY (SEQ ID NO:314) ADSVKGRFTISRDNPKNTLYLQMNSLRAEDTAVYYCARGGGIAVADYYFYGMDVWGQGTT VTVSSL15.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 315)ATCTCCTGCAGGTCTAGTCAAAGCCTCATATACAGTGATGGAAACACTTACTTGAATTGGTTTCAACAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTAACTGGGACTCTGGGGTCCCAGACAGATTCAGCGGCAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGATTTATTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L15.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLIYSDGNTYLNWFQQRPGQSPRRLIYKVSNWDSGV (SEQ IDNO: 316) PDRFSGSGSGTDFTLKISRVEAEDVGIYYCMQGTHWPPAFGQGTRLEIK L15.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:317) ATTACTTGTCGGGCGAGTCAGGGTATTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGGTCCTGACCTATACTACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCTACTTACTTTTGTCAACAGGGTGACAGTTTCCCTCTCACTTTTCGGCGGGGGGACCAAGGTGGAGATCAAA L15.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGISSWLAWYQQKPGKAPKVLTYTTSSLQSGVPSRFS (SEQ IDNO: 318) GSGSGTDFTLTISSLQPEDFATYFCQQADSFPLTFGGGTKVEIK H16 DNACAGGTGCAACTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:319) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATAGCAGTGGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTGCTCA H16 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVIWYDGSNKY (SEQ ID NO:320) YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIAVADYYYYGMDVWGQG TTVTVSSL16.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 321)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTATACAGTGATGGAAACACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTTACTGGGACTCTGGGGTCCCAGACAGATTCAGCGGCAGTGGGTCAAGCACTGATTTCACACTGAAAATCAGTAGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L16.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGNTYLNWFQQRPGQSPRRLIYKVSYWDSG (SEQ IDNO: 322) VPDRFSGSGSSTDFTLKISRVEAEDVGVYYCMQGTHWPPAFGQGTRLEIK L16.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:323) ATCACTTGTCGGGCGAGTCAGAGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGCTCCATAATGCATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGTAAATTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAGGGTGGAGATCAAA L16.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQSLSSWLAWYQQKPGKAPKLLLHNASSLQSGVPSRFS (SEQ IDNO: 324) GSGSGTDFTLTISSLQPEDFVNYYCQQANSFPLTFGGGTRVEIK H17 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTAAGACT (SEQ ID NO:325) CTCCTGTGCAGCGTCTGGATTCACCTTAAGTAGTTATGGCATGCTCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTTTATGGTTTGATGGAAGTTATAAAAACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGCGAGCGGAGGACACGGCTGTGTATTACTGTGCGAGAGATAGTACAACTATGGCCCACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTC A H17Protein QVQLVESGGGVVQPGRSLRLSCAASGFTLSSYGMLWVRQAPGKGLEWVAVLWFDGSYKNY(SEQ ID NO: 326)ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSTTMAHFDYWGQGTLVTVSS L17 DNACAGACTGTGGTGACCCAGGAGCCATCGTTCTCAGTGTCCCCTGGAGGGACAGTCACACTC (SEQ ID NO:327) ACTTGTGGCTTGAACTCTGGCTCAGTCTCTACTAGTTACTTCCCCAGCTGGTACCAGCAGACCCCAGGCCAGGCTCCACGCACGCTCATCTACAGCACAAACAGTCGCTCTTCTGGGGTCCCTGATCGCTTCTCTGGCTCCATCCTTGGGAACAAAGCTGCCCTCACCATCACGGGGGCCCAGGCAGATGATGAATCTGATTATTACTGTGTGCTGTATATGGGTAGAGGCATTTGGGTGTTTCGGCGGAGGGACCAAGCTGACCGTCCTA L17 ProteinQTVVTQEPSFSVSPGGTVTLTCGLNSGSVSTSYFPSWYQQTPGQAPRTLIYSTNSRSSGVPDRF (SEQ IDNO: 328) SGSILGNKAALTITGAQADDESDYYCVLYMGRGIWVFGGGTKLTVL H18 DNACAGGTGCAACTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:319) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAACTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATAGCAGTGGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H18 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEWVAVIWYDGSNKY (SEQ ID NO:320) YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIAVADYYYYGMDVWGQG TTVTVSSL18.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 329)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTATACAGTGATGGAAACACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTTACTGGGACTCTGGGGTCCCAGACAGTTCAGCGGCAGTGGGTCAGGCACTGATTTCACACTGAAAATCAGTAGGGTGGAGGCTGAGGATGTTGGGGTTTATTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATCAAA L18.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGNTYLNWFQQRPGQSPRRLIYKVSYWDSG (SEQ IDNO: 330) VPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQGTHWPPAFGQGTRLEIK L18.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:331) ATCACTTGTCGGGCGAGTCAGAGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGCTCTATAATGCATCCAGTTTGCAAAGTGGGGCCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGTAACTTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAGGGTGGAGATCAAA L18.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQSLSSWLAWYQQKPGKAPKLLLYNASSLQSGAPSRFS (SEQ IDNO: 332) GSGSGTDFTLTISSLQPEDFVTYYCQQANSFPLTFGGGTRVEIK H19 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:304) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATACCAGTAGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H19 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:305) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIPVADYYYYGMDVWGQGTT VTVSSL19.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 306)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTCTACAGTGATGGAGACACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTAACTGGGACTCTGGGGTCCCATACAGATTCAGCGGCAGTGGGTCAGGCACTGATTTCACACTGCAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGATTTACTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L19.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGDTYLNWFQQRPGQSPRRLIYKVSNWDSG (SEQ IDNO: 307) VPYRFSGSGSGTDFTLQISRVEAEDVGIYYCMQGTHWPPAFGQGTRLEIK L19.2 DNAGACATCCAGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:308) ATCACTTGTCGGGCGAGTCAGGGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCCTGATGTATAACACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAGTTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA L19.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGLSSWLAWYQQKPGKAPKLLMYNTSSLQSGVPSRF (SEQ IDNO: 309) SGSGSGTDFSLTISSLQPEDFASYYCQQANSFPLTFGGGTKVEIK H20 DNACAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:304) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGGGGGGGGTATACCAGTAGCTGACTACTACTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H20 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:305) ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARGGGIPVADYYYYGMDVWGQGTT VTVSSL20.1 DNA GATGTTGTGATGACTCAGTCTCCACTCTCCCTGCCCGTCACCCTTGGACAGCCGGCCTCC(SEQ ID NO: 306)ATCTCCTGCAGGTCTAGTCAAAGCCTCGTCTACAGTGATGGAGAGACCTACTTGAATTGGTTTCAGCAGAGGCCAGGCCAATCTCCAAGGCGCCTAATTTATAAGGTTTCTAACTGGGACTCTGGGGTCCCATACAGATTCAGCGGCAGTGGGTCAGGCACTGATTTCACACTGCAAATCAGCAGGGTGGAGGCTGAGGATGTTGGGATTTACTACTGCATGCAAGGTACACACTGGCCTCCGGCCTTTCGGCCAAGGGACACGACTGGAGATTAAA L20.1 ProteinDVVMTQSPLSLPVTLGQPASISCRSSQSLVYSDGDTYLNWFQQRPGQSPRRLIYKVSNWDSG (SEQ IDNO: 307) VPYRFSGSGSGTDFTLQISRVEAEDVGIYYCMQGTHWPPAFGQGTRLEIK L20.2 DNAGACATCCAGATGACCCAGTCCCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACC (SEQ ID NO:333) ATCACTTGTCGGGCGAGTCAGGGTCTTAGCAGCTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCCAAGCTCCTGATGTATAACACATCCAGTTTGCAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCAGTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAAGTTACTATTGTCAACAGGCTAACAGTTTCCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA L20.2 ProteinDIQMTQSPSSVSASVGDRVTITCRASQGLSSWLAWYQQKPGKAPKLLMYNTSSLQSGVPSRF (SEQ IDNO: 309) SGSGSGTDFSLTISSLQPEDFASYYCQQANSFPLTFGGGTKVEIK H21 DNAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACT (SEQ ID NO:334) CTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCAATTAGTGGTAGTGGTGGAAGTACACACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAGATCTCAACTGGGGAGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTCA H21 ProteinEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTHYA (SEQ IDNO: 335) DSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDLNWGAFDIWGQGTMVTVSS L21DNA CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTGGGGCCCCAGGGCAGAGGGTCACCAT (SEQ IDNO: 336) CTCCTGCACTGGGAGCAGCTCCAACATTGGGGCGGGTTATGTTGTACATTGGTACCAGCAGCTTCCAGGAACAGCCCCCAAACTCCTCATCTATGGTAACAGCAATCGGCCCTCAGGGGTCCCTGACCAATTCTCTGGCTCCAAGTCTGGCACCTCAGCCTCCCTGGCCATCACTGGACTCCAGTCTGAGGATGAGGCTGATTATTACTGCAAAGCATGGGATAACAGCCTGAATGCTCAAGGGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L21 ProteinQSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYVVHWYQQLPGTAPKLLIYGNSNRPSGVPDQ (SEQ IDNO: 337) FSGSKSGTSASLAITGLQSEDEADYYCKAWDNSLNAQGVFGGGTKLTVL H22 DNAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGCACAGCCGGGGGGGTCCCTGAGACT (SEQ ID NO:338) CTCCTGTGGAGGCTCTGGATTCTCCTTTAGAGGCTATGTCATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGAATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTGTCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAGGAGACAGCTCGAACTACTACTCCGGTATGGACGTCTGGGGCCAAGGGACCACGGTCATCGT CTCCTCA H22Protein EVQLLESGGGLAQPGGSLRLSCAGSGFSFRGYVMTWVRQAPGKGLEWVSGISGSGGSTYYA(SEQ ID NO: 339)DSVKGRFTISRDNSKNTLCLQMNSLRAEDTAVYYCAKGDSSNYYSGMDVWGQGTTVIVSS L22 DNAGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACC (SEQ ID NO:340) ATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAACTCCAACAATAAGAACTACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCTTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAGGATGTGGCAATTTATTACTGTCAGCAATTTTATGGTCCTCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAAATCAAA L22 ProteinDIVMTQSPDSLAVSLGERATINCKSSQSVLYNSNNKNYLAWYQQKPGQPPKLLIYWASTRES (SEQ IDNO: 341) GVPDRFSGSGSGTDFTLTISSLQAEDVAIYYCQQFYGPPLTFGGGTKVEIK H23 DNACAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAGTGAAGGT (SEQ ID NO:342) CTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCACTGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACCCTAACAATGGTGGCACAAACTATGGACAGAAGTTTCAGGGCAGGGTCACCATGACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGATCTGACGACACGGCCGTGTATTACTGTGCGAGAGGGAACTGGAACGACGATGCTTTTGATATCTGGGGCCAAGGGACAATGGTCACCGTCTCTTC A H23Protein QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNNGGTN (SEQID NO: 343) YGQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGNWNDDAFDIWGQGTMVTVSSL23 DNA TCCTATGAGCTGACTCAGTCACCCTCAGTGTCCGTGTCCCCAGGACAGACAGCCAGCATC(SEQ ID NO: 344)ACCTGTTCTGGTGATAAATTGGGGGATAAATTTGCTTTCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTGCTGGTCATCTATCAAGATAGCAAGCGGCCCTCAGGGATCCCTGAGCGATTCTCTGGCTCCAACTCTGGGAACACAGCCACTCTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGCAGCGCCGGGGGGGTATTTCGGCGGAGGGACCAAGTTGACCGTCCTA L23 ProteinSYELTQSPSVSVSPGQTASITCSGDKLGDKFAFWYQQKPGQSPVLVIYQDSKRPSGIPERFSGS (SEQ IDNO: 345) NSGNTATLTISGTQAMDEADYYCQAWDSSAGGVFGGGTKLTVL H24 DNACAGGTGCAACTGGAGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:346) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTATATGGTATGATGGAAGTAATAAATACTATGTAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAATGGGGTTTACTATGGTTCGGGGAGCCCTCTACTAGGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H24 ProteinQVQLEESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVIWYDGSNKYY (SEQ ID NO:347) VDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARMGFTMYRGALYYGMDVWGQGT TVTVSSL24 DNA TCTTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATC(SEQ ID NO: 348)ACATGCCAAGGAGACAGCCTCAGAAGCTATCATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTGAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGACTCCAGTTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTATTGTAATTATCGGGACAACAGTGGTAACCATCTGGTGTTTCGGCGGAGGGACCAAGCTGACCGTCCTA L24 ProteinSSELTQDPAVSVALGQTVRITCQGDSLRSYHASWYQQKPGQAPVLVIYGENNRPSGIPDRFSD (SEQ IDNO: 349) SSSGNTASLTITGAQAEDEADYYCNYRDNSGNHLVFGGGTKLTVL H25 DNAGAGGTGCAGCTGTTGGAATCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACT (SEQ ID NO:350) CTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGGCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTCGTAGTGGTAGTACCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGTGGAACCGAGATATTTTGACTGGTTATTAGGCGACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTC A H25Protein EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISRSGSTTYYAD(SEQ ID NO: 351)SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVEPRYFDWLLGDWGQGTLVTVSS L25 DNAGACATCGTGATGACCCAGTCTCCAGACTCCCTGGCTGTGTCTCTGGGCGAGAGGGCCACC (SEQ ID NO:340) ATCAACTGCAAGTCCAGCCAGAGTGTTTTATACAACTCCAACAATAAGAACTACTTAGCTTGGTACCAGCAGAAACCAGGACAGCCTCCTAAGCTGCTCATTTACTGGGCTTCTACCCGGGAATCCGGGGTCCCTGACCGATTCAGTGGCAGCGGGTCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGGCTGAGGATGTGGCAATTTATTACTGTCAGCAATTTTATGGTCCTCCTCTCACTTTTCGGCGGAGGGACCAAGGTGGAAATCAAA L25 ProteinDIVMTQSPDSLAVSLGERATINCKSSQSVLYNSNNKNYLAWYQQKPGQPPKLLIYWASTRES (SEQ IDNO: 341) GVPDRFSGSGSGTDFTLTISSLQAEDVAIYYCQQFYGPPLTFGGGTKVEIK H26 DNACAGGTGCAGCTGGTGGAGTCGGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACT (SEQ ID NO:352) CTCCTGTGCAGCGTCTGGATTCACCTTCAGTAGCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTGGGTGGCAGTTAAATGGTATGAAGGAAGTAATAAATACTATGGAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATTTGCAAATGAACAGTCTGAGAGGCGAGGATACGGCTGTGTATTACTGTGCGAGAGGCGCCCACGACTACGGTGACTTCTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCA H26 ProteinQVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEWVAVKWYEGSNKY (SEQ ID NO:353) YGDSVKGRFTISRDNSKNTLYLQMNSLRGEDTAVYYCARGAHDYGDFYYGMDVWGQGTT VTVSSL26 DNA TCCTATGAACTGACTCAGCCAGCCTCAGTGTCCGTGTCCCCAGGACAGATAGCCAGCATC(SEQ ID NO: 354)ACCTGCTCTGGAGATAATTTGGGGGATAAATATATTTGCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTGCGGGTCATCTATCAAGATAACAAGCGGCCCTCAGGGATCCCTGAGCGTTTCTCTGGCTCCAATTCTGGGAACACAGCCACTCTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGCAGCACTGTGGTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L26 ProteinSYELTQPASVSVSPGQIASITCSGDNLGDKYICWYQQKPGQSPVRVIYQDNKRPSGIPERFSGS (SEQ IDNO: 355) NSGNTATLTISGTQAMDEADYYCQAWDSSTVVFGGGTKLTVL H27 DNAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACT (SEQ ID NO:356) CTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTTATAGTGGCGGTAGCACATACTACGCAGGCTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAGATCGGGAGGGAGCGACTTGGTACTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCA CCGTCTCCTCAH27 ProteinEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISYSGGSTYYA (SEQ IDNO: 357) GSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKDREGATWYYGMDVWGQGTTVTV SSL27 DNA TCCTATGAACTGACTCAGCCACCCTCAGTGTCCGTGTCCCCAGGACAGACAGCCAGCATC(SEQ ID NO: 358)ACCTGCTCTGGAGATAAATTGGGGGAAAGCTATGCTTGCTGGTATCAGCAGAAGCCAGGCCAGTCCCCTGTACTGGTCATCTATCAAGATTACAAGCGGCCCTCAGGGATCCCTGAGCGCTTCTCTGGCTCCAACTCTGGGAACACAGCCACTCTGACCATCAGCGGGACCCAGGCTATGGATGAGGCTGACTATTACTGTCAGGCGTGGGACAGAAGTACTGTACTATTTCGGCGGAGGGACCAAGCTGACCGTCCTA L27 ProteinSYELTQPPSVSVSPGQTASITCSGDKLGESYACWYQQKPGQSPVLVIYQDYKRPSGIPERFSGS (SEQ IDNO: 359) NSGNTATLTISGTQAMDEADYYCQAWDRSTVLFGGGTKLTVL

Particular embodiments of antigen binding proteins of the presentinvention comprise one or more amino acid sequences that are identicalto the amino acid sequences of one or more of the CDRs and may furthercomprise one or more FRs illustrated above. In one embodiment, theantigen binding protein comprises a light chain CDR1 sequenceillustrated above. In another embodiment, the antigen binding proteincomprises a light chain CDR2 sequence illustrated above. In anotherembodiment, the antigen binding protein comprises a light chain CDR3sequence illustrated above. In another embodiment, the antigen bindingprotein comprises a heavy chain CDR1 sequence illustrated in above. Inanother embodiment, the antigen binding protein comprises a heavy chainCDR2 sequence illustrated above. In another embodiment, the antigenbinding protein comprises a heavy chain CDR3 sequence illustrated above.In another embodiment, the antigen binding protein further comprises alight chain FR1 sequence illustrated above. In another embodiment, theantigen binding protein further comprises a light chain FR2 sequenceillustrated above. In another embodiment, the antigen binding proteinfurther comprises a light chain FR3 sequence illustrated above. Inanother embodiment, the antigen binding protein further comprises alight chain FR4 sequence illustrated above. In another embodiment, theantigen binding protein further comprises a heavy chain FR1 sequenceillustrated above. In another embodiment, the antigen binding proteinfurther comprises a heavy chain FR2 sequence illustrated above. Inanother embodiment, the antigen binding protein further comprises aheavy chain FR3 sequence illustrated above. In another embodiment, theantigen binding protein further comprises a heavy chain FR4 sequenceillustrated above.

In one embodiment, the present disclosure provides an antigen bindingprotein comprising a light chain variable domain comprising a sequenceof amino acids that differs from the sequence of a light chain variabledomain selected from the group consisting of L1 through L27 only at 15,14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 residues, whereineach such sequence difference is independently either a deletion,insertion, or substitution of one amino acid residue. In anotherembodiment, the light-chain variable domain comprises a sequence ofamino acids that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99%identical to the sequence of a light chain variable domain selected fromthe group consisting of L1-L27. In another embodiment, the light chainvariable domain comprises a sequence of amino acids that is encoded by anucleotide sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%,or 99% identical to a nucleotide sequence that encodes a light chainvariable domain selected from the group consisting of L1-L27. In anotherembodiment, the light chain variable domain comprises a sequence ofamino acids that is encoded by a polynucleotide that hybridizes undermoderately stringent conditions to the complement of a polynucleotidethat encodes a light chain variable domain selected from the groupconsisting of L1-L27. In another embodiment, the light chain variabledomain comprises a sequence of amino acids that is encoded by apolynucleotide that hybridizes under highly stringent conditions to acomplement of a light chain polynucleotide of L1-L27.

In another embodiment, the present invention provides an antigen bindingprotein comprising a heavy chain variable domain comprising a sequenceof amino acids that differs from the sequence of a heavy chain variabledomain selected from the group consisting of H1-H27 only at 15, 14, 13,12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or 0 residue(s), wherein each suchsequence difference is independently either a deletion, insertion, orsubstitution of one amino acid residue. In another embodiment, the heavychain variable domain comprises a sequence of amino acids that is atleast 70%, 75%, 80%, 85%, 90%, 95%, 97%, or 99% identical to thesequence of a heavy chain variable domain selected from the groupconsisting of H1-H27. In another embodiment, the heavy chain variabledomain comprises a sequence of amino acids that is encoded by anucleotide sequence that is at least 70%, 75%, 80%, 85%, 90%, 95%, 97%,or 99% identical to a nucleotide sequence that encodes a heavy chainvariable domain selected from the group consisting of H1-H27. In anotherembodiment, the heavy chain variable domain comprises a sequence ofamino acids that is encoded by a polynucleotide that hybridizes undermoderately stringent conditions to the complement of a polynucleotidethat encodes a heavy chain variable domain selected from the groupconsisting of H1-H27. In another embodiment, the heavy chain variabledomain comprises a sequence of amino acids that is encoded by apolynucleotide that hybridizes under highly stringent conditions to thecomplement of a polynucleotide that encodes a heavy chain variabledomain selected from the group consisting of H1-H27.

In some of the embodiments provided in Table 2 above, two light chainsare associated with a single heavy chain, identified, for example asL-12.1 μL-12.2, etc. These alternative light chains are each paired witha single heavy chain. In these embodiments, light chain and heavy chaincombination may be assayed as described below and the combination oflight chain and heavy chain that provides the greater TSLP neutralizingactivity may be selected.

Additional embodiments include antigen binding proteins comprising thecombinations L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9,L10H10, L11H11, L12H12, L13H13, L14H14, L15H15, L16H16, L17H17, L18H18,L19H19, L20H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26, andL27H27.

Antigen binding proteins (e.g., antibodies, antibody fragments, andantibody derivatives) of the invention can further comprise any constantregion known in the art. The light chain constant region can be, forexample, a kappa- or lambda-type light chain constant region, e.g., ahuman kappa- or lambda-type light chain constant region. The heavy chainconstant region can be, for example, an alpha-, delta-, epsilon-,gamma-, or mu-type heavy chain constant regions, e.g., a human alpha-,delta-, epsilon-, gamma-, or mu-type heavy chain constant region. In oneembodiment, the light or heavy chain constant region is a fragment,derivative, variant, or mutein of a naturally occurring constant region.

In one embodiment, the antigen binding proteins comprise an IgG, such asIgG1, IgG2, IgG3, or IgG4.

Techniques are known for deriving an antibody of a different subclass orisotype from an antibody of interest, i.e., subclass switching. Thus,IgG antibodies may be derived from an IgM antibody, for example, andvice versa. Such techniques allow the preparation of new antibodies thatpossess the antigen-binding properties of a given antibody (the parentantibody), but also exhibit biological properties associated with anantibody isotype or subclass different from that of the parent antibody.Recombinant DNA techniques may be employed. Cloned DNA encodingparticular antibody polypeptides may be employed in such procedures,e.g., DNA encoding the constant domain of an antibody of the desiredisotype. See also Lantto et al., 2002, Methods Mol. Biol. 178:303-16.

In one embodiment, an antigen binding protein of the invention comprisesthe IgG1 heavy chain constant domain or a fragment of the IgG1 heavychain domain. In one embodiment, an antigen binding protein of theinvention further comprises the constant light chain kappa or lambdadomains or a fragment of these. Light chain constant regions andpolynucleotides encoding them are provided in Table 3 below. In anotherembodiment, an antigen binding protein of the invention furthercomprises a heavy chain constant domain, or a fragment thereof, such asthe IgG2 heavy chain constant region shown below in Table 3.

The nucleic acid (DNA) encoding constant heavy and constant light chaindomains, and the amino acids sequences of heavy and light chain domainsare provided below. Lambda variable domains can be fused to lambdaconstant domains and kappa variable domains can be fused to kappaconstant domains.

TABLE 3 (SEQ ID NO: 364) IgG2 Heavy Constant domain DNAgctagcaccaagggcccatcggtcttccccctggcgccctgctccaggagcacctccgagagcacagcggccctgggctgcctggtcaaggactacttccccgaaccggtgacggtgtcgtggaactcaggcgctctgaccagcggcgtgcacaccttcccagctgtcctacagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcaacttcggcacccagacctacacctgcaacgtagatcacaagcccagcaacaccaaggtggacaagacagttgagcgcaaatgttgtgtcgagtgcccaccgtgcccagcaccacctgtggcaggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctgaggtcacgtgcgtggtggtggacgtgagccacgaagaccccgaggtccagttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccacgggaggagcagttcaacagcacgttccgtgtggtcagcgtcctcaccgttgtgcaccaggactggctgaacggcaaggagtacaagtgcaaggtctccaacaaaggcctcccagcccccatcgagaaaaccatctccaaaaccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctaccccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacacctcccatgctggactccgacggctccttcttcctctacagcaagctcaccgtggacaagagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccactacacgcagaagagcctctccctgtctccgggtaaatga (SEQ ID NO: 365) IgG2 HeavyConstant domain ProteinASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTFPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK* (SEQ ID NO: 366) Kappa Light Constant domain DNAcagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaacaggggagagtgttag (SEQ ID NO: 367) Kappa LightConstant domain ProteinRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* (SEQ ID NO: 368) LambdaLight Constant domain DNAggccaaccgaaagcggcgccctcggtcactctgttcccgccctcctctgaggagcttcaagccaacaaggccacactggtgtgtctcataagtgacttctacccgggagccgtgacagtggcctggaaggcagatagcagccccgtcaaggcgggagtggagaccaccacaccctccaaacaaagcaacaacaagtacgcggccagcagctatctgagcctgacgcctgagcagtggaagtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacagaatgttcatag (SEQ ID NO: 369) Lambda LightConstant domain ProteinGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS*

The antigen binding proteins of the present invention include thosecomprising, for example, the variable domain combinations L1H1, L2H2,L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12,L13.1H13, L13.2H13, L14.1H14, L14.2H14, L15.1H15, L15.2H15, L16.1H16,L16.2H16, L17H17, L18.1H18, L18.2H18, L19.1H19, L19.2H19, L20.1H20,L20.2H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26, and L27H27.having a desired isotype (for example, IgA, IgG1, IgG2, IgG3, IgG4, IgM,IgE, and IgD) as well as Fab or F(ab′)₂ fragments thereof. Moreover, ifan IgG4 is desired, it may also be desired to introduce a point mutationin the hinge region as described in Bloom et al., 1997, Protein Science6:407 (incorporated by reference herein) to alleviate a tendency to formintra-H chain disulfide bonds that can lead to heterogeneity in the IgG4antibodies.

Antibodies and Antibody Fragments

As used herein, the term “antibody” refers to an intact antibody, or anantigen binding fragment thereof, as described in the definition sectionherein. An antibody may comprise a complete antibody molecule (includingpolyclonal, monoclonal, chimeric, humanized, or human versions havingfull length heavy and/or light chains), or comprise an antigen bindingfragment thereof. Antibody fragments include F(ab′)₂, Fab, Fab′, Fv, Fc,and Fd fragments, and can be incorporated into single domain antibodies,monovalent antibodies, single-chain antibodies, maxibodies, minibodies,intrabodies, diabodies, triabodies, tetrabodies, v-NAR and bis-scFv (Seee.g., Hollinger and Hudson, 2005, Nature Biotechnology, 23, 9,1126-1136). Antibody polypeptides are also disclosed in U.S. Pat. No.6,703,199, including fibronectin polypeptide monobodies. Other antibodypolypeptides are disclosed in U.S. Patent Publication 2005/0238646,which are single-chain polypeptides. Monovalent antibody fragments aredisclosed in US Patent Publication 20050227324.

Antigen binding fragments derived from an antibody can be obtained, forexample, by proteolytic hydrolysis of the antibody, for example, pepsinor papain digestion of whole antibodies according to conventionalmethods. By way of example, antibody fragments can be produced byenzymatic cleavage of antibodies with pepsin to provide a 5S fragmenttermed F(ab′)₂. This fragment can be further cleaved using a thiolreducing agent to produce 3.5S Fab′ monovalent fragments. Optionally,the cleavage reaction can be performed using a blocking group for thesulfhydryl groups that result from cleavage of disulfide linkages. As analternative, an enzymatic cleavage using papain produces two monovalentFab fragments and an Fc fragment directly. These methods are described,for example, by Goldenberg, U.S. Pat. No. 4,331,647, Nisonoff et al.,Arch. Biochem. Biophys. 89:230, 1960; Porter, Biochem. J. 73:119, 1959;Edelman et al., in Methods in Enzymology 1:422 (Academic Press 1967);and by Andrews, S. M. and Titus, J. A. in Current Protocols inImmunology (Coligan J. E., et al., eds), John Wiley & Sons, New York(2003), pages 2.8.1-2.8.10 and 2.10A. 1-2.10A.5. Other methods forcleaving antibodies, such as separating heavy chains to form monovalentlight-heavy chain fragments (Fd), further cleaving of fragments, orother enzymatic, chemical, or genetic techniques may also be used, solong as the fragments bind to the antigen that is recognized by theintact antibody.

An antibody fragment may also be any synthetic or genetically engineeredprotein. For example, antibody fragments include isolated fragmentsconsisting of the light chain variable region, “Fv” fragments consistingof the variable regions of the heavy and light chains, recombinantsingle chain polypeptide molecules in which light and heavy variableregions are connected by a peptide linker (scFv proteins).

Another form of an antibody fragment is a peptide comprising one or morecomplementarity determining regions (CDRs) of an antibody. CDRs (alsotermed “minimal recognition units”, or “hypervariable region”) can beobtained by constructing polynucleotides that encode the CDR ofinterest. Such polynucleotides are prepared, for example, by using thepolymerase chain reaction to synthesize the variable region using mRNAof antibody-producing cells as a template (see, for example, Larrick etal., Methods: A Companion to Methods in Enzymology 2:106, 1991;Courtenay-Luck, “Genetic Manipulation of Monoclonal Antibodies,” inMonoclonal Antibodies: Production, Engineering and Clinical Application,Ritter et al. (eds.), page 166 (Cambridge University Press 1995); andWard et al., “Genetic Manipulation and Expression of Antibodies,” inMonoclonal Antibodies: Principles and Applications, Birch et al.,(eds.), page 137 (Wiley-Liss, Inc. 1995)).

Thus, in one embodiment, the binding agent comprises at least one CDR asdescribed herein. The binding agent may comprise at least two, three,four, five or six CDR's as described herein. The binding agent furthermay comprise at least one variable region domain of an antibodydescribed herein. The variable region domain may be of any size or aminoacid composition and will generally comprise at least one CDR sequenceresponsible for binding to TSLP, for example heavy chain CDR1, CDR2,CDR3 and/or the light chain CDRs specifically described herein and whichis adjacent to or in frame with one or more framework sequences. Ingeneral terms, the variable (V) region domain may be any suitablearrangement of immunoglobulin heavy (V_(H)) and/or light (V_(L)) chainvariable domains. Thus, for example, the V region domain may bemonomeric and be a V_(H) or V_(L) domain, which is capable ofindependently binding human TSLP with an affinity at least equal to1×10⁻⁷M or less as described below. Alternatively, the V region domainmay be dimeric and contain V_(H)-V_(H), V_(H)-V_(L), or V_(L)-V_(L),dimers. The V region dimer comprises at least one V_(H) and at least oneV_(L) chain that may be non-covalently associated (hereinafter referredto as F_(V)). If desired, the chains may be covalently coupled eitherdirectly, for example via a disulfide bond between the two variabledomains, or through a linker, for example a peptide linker, to form asingle chain Fv (scF_(V)).

The variable region domain may be any naturally occurring variabledomain or an engineered version thereof. By engineered version is meanta variable region domain that has been created using recombinant DNAengineering techniques. Such engineered versions include those created,for example, from a specific antibody variable region by insertions,deletions, or changes in or to the amino acid sequences of the specificantibody. Particular examples include engineered variable region domainscontaining at least one CDR and optionally one or more framework aminoacids from a first antibody and the remainder of the variable regiondomain from a second antibody.

The variable region domain may be covalently attached at a C-terminalamino acid to at least one other antibody domain or a fragment thereof.Thus, for example, a VH domain that is present in the variable regiondomain may be linked to an immunoglobulin CH1 domain, or a fragmentthereof. Similarly a V_(L) domain may be linked to a C_(K) domain or afragment thereof. In this way, for example, the antibody may be a Fabfragment wherein the antigen binding domain contains associated V_(H)and V_(L) domains covalently linked at their C-termini to a CH1 andC_(K) domain, respectively. The CH1 domain may be extended with furtheramino acids, for example to provide a hinge region or a portion of ahinge region domain as found in a Fab′ fragment, or to provide furtherdomains, such as antibody CH2 and CH3 domains.

Derivatives of Antigen Binding Proteins

The nucleotide sequences shown in FIG. 1A-1F, FIG. 2A-2F, and Table 2above can be altered, for example, by random mutagenesis or bysite-directed mutagenesis (e.g., oligonucleotide-directed site-specificmutagenesis) to create an altered polynucleotide comprising one or moreparticular nucleotide substitutions, deletions, or insertions ascompared to the non-mutated polynucleotide. Examples of techniques formaking such alterations are described in Walder et al., 1986, Gene42:133; Bauer et al. 1985, Gene 37:73; Craik, BioTechniques, Jan. 1985,12-19; Smith et al., 1981, Genetic Engineering: Principles and Methods,Plenum Press; and U.S. Pat. Nos. 4,518,584 and 4,737,462. These andother methods can be used to make, for example, derivatives of TSLPantigen binding proteins that have a desired property, for example,increased affinity, avidity, or specificity for TSLP, increased activityor stability in vivo or in vitro, or reduced in vivo side-effects ascompared to the underivatized antigen binding proteins.

Other derivatives of anti-TSLP antigen binding proteins includingantibodies within the scope of this invention include covalent oraggregative conjugates of anti-TSLP antibodies, or fragments thereof,with other proteins or polypeptides, such as by expression ofrecombinant fusion proteins comprising heterologous polypeptides fusedto the N-terminus or C-terminus of an anti-TSLP antibody polypeptide.For example, the conjugated peptide may be a heterologous signal (orleader) polypeptide, e.g., the yeast alpha-factor leader, or a peptidesuch as an epitope tag. Antigen binding protein-containing fusionproteins can comprise peptides added to facilitate purification oridentification of antigen binding protein (e.g., poly-His). An antigenbinding protein also can be linked to the FLAG peptide as described inHopp et al., Bio/Technology 6:1204, 1988, and U.S. Pat. No. 5,011,912.The FLAG peptide is highly antigenic and provides an epitope reversiblybound by a specific monoclonal antibody (mAb), enabling rapid assay andfacile purification of expressed recombinant protein. Reagents usefulfor preparing fusion proteins in which the FLAG peptide is fused to agiven polypeptide are commercially available (Sigma, St. Louis, Mo.).

Oligomers that contain one or more antigen binding proteins may beemployed as TSLP antagonists. Oligomers may be in the form ofcovalently-linked or non-covalently-linked dimers, trimers, or higheroligomers. Oligomers comprising two or more antigen binding proteins arecontemplated for use, with one example being a homodimer. Otheroligomers include heterodimers, homotrimers, heterotrimers,homotetramers, heterotetramers, etc.

One embodiment is directed to oligomers comprising multiple antigenbinding proteins joined via covalent or non-covalent interactionsbetween peptide moieties fused to the antigen binding proteins. Suchpeptides may be peptide linkers (spacers), or peptides that have theproperty of promoting oligomerization. Leucine zippers and certainpolypeptides derived from antibodies are among the peptides that canpromote oligomerization of antigen binding proteins attached thereto, asdescribed in more detail below.

In particular embodiments, the oligomers comprise from two to fourantigen binding proteins capable of binding to TSLP. The antigen bindingproteins of the oligomer may be in any form, such as any of the formsdescribed above, e.g., variants or fragments.

In one embodiment, an oligomer is prepared using polypeptides derivedfrom immunoglobulins. Preparation of fusion proteins comprising certainheterologous polypeptides fused to various portions of antibody-derivedpolypeptides (including the Fc domain) has been described, e.g., byAshkenazi et al., 1991, PNAS USA 88:10535; Byrn et al., 1990, Nature344:677; and Hollenbaugh et al., 1992 “Construction of ImmunoglobulinFusion Proteins”, in Current Protocols in Immunology, Suppl. 4, pages10.19.1-10.19.11.

One embodiment of the present invention is directed to a dimercomprising two fusion proteins created by fusing a fragment of ananti-TSLP antibody to the Fc region of an antibody. The dimer can bemade by, for example, inserting a gene fusion encoding the fusionprotein into an appropriate expression vector, expressing the genefusion in host cells transformed with the recombinant expression vector,and allowing the expressed fusion protein to assemble much like antibodymolecules, whereupon interchain disulfide bonds form between the Fcmoieties to yield the dimer.

The term “Fc polypeptide” as used herein includes native and muteinforms of polypeptides derived from the Fc region of an antibody.Truncated forms of such polypeptides containing the hinge region thatpromotes dimerization also are included. Fusion proteins comprising Fcmoieties (and oligomers formed therefrom) offer the advantage of facilepurification by affinity chromatography over Protein A or Protein Gcolumns.

One suitable Fc polypeptide, described in PCT application WO 93/10151(hereby incorporated by reference), is a single chain polypeptideextending from the N-terminal hinge region to the native C-terminus ofthe Fc region of a human IgG1 antibody. Another useful Fc polypeptide isthe Fc mutein described in U.S. Pat. No. 5,457,035 and in Baum et al.,1994, EMBO J. 13:3992-4001. The amino acid sequence of this mutein isidentical to that of the native Fc sequence presented in WO 93/10151,except that amino acid 19 has been changed from Leu to Ala, amino acid20 has been changed from Leu to Glu, and amino acid 22 has been changedfrom Gly to Ala. The mutein exhibits reduced affinity for Fc receptors.

In other embodiments, the variable portion of the heavy and/or lightchains of an anti-TSLP antibody may be substituted for the variableportion of an antibody heavy and/or light chain.

Alternatively, the oligomer is a fusion protein comprising multipleantigen binding proteins, with or without peptide linkers (spacerpeptides). Among the suitable peptide linkers are those described inU.S. Pat. Nos. 4,751,180 and 4,935,233.

Another method for preparing oligomeric antigen binding proteinsinvolves use of a leucine zipper. Leucine zipper domains are peptidesthat promote oligomerization of the proteins in which they are found.Leucine zippers were originally identified in several DNA-bindingproteins (Landschulz et al., 1988, Science 240:1759), and have sincebeen found in a variety of different proteins. Among the known leucinezippers are naturally occurring peptides and derivatives thereof thatdimerize or trimerize. Examples of leucine zipper domains suitable forproducing soluble oligomeric proteins are described in PCT applicationWO 94/10308, and the leucine zipper derived from lung surfactant proteinD (SPD) described in Hoppe et al., 1994, FEBS Letters 344:191, herebyincorporated by reference. The use of a modified leucine zipper thatallows for stable trimerization of a heterologous protein fused theretois described in Fanslow et al., 1994, Semin. Immunol. 6:267-78. In oneapproach, recombinant fusion proteins comprising an anti-TSLP antibodyfragment or derivative fused to a leucine zipper peptide are expressedin suitable host cells, and the soluble oligomeric anti-TSLP antibodyfragments or derivatives that form are recovered from the culturesupernatant.

As described herein, antibodies comprise at least one CDR. For example,one or more CDR may be incorporated into known antibody frameworkregions (IgG1, IgG2, etc.), or conjugated to a suitable vehicle toenhance the half-life thereof. Suitable vehicles include, but are notlimited to Fc, polyethylene glycol (PEG), albumin, transferrin, and thelike. These and other suitable vehicles are known in the art. Suchconjugated CDR peptides may be in monomeric, dimeric, tetrameric, orother form. In one embodiment, one or more water-soluble polymer isbonded at one or more specific position, for example at the aminoterminus, of a binding agent.

In certain preferred embodiments, an antibody comprises one or morewater soluble polymer attachments, including, but not limited to,polyethylene glycol, polyoxyethylene glycol, or polypropylene glycol.See, e.g., U.S. Pat. Nos. 4,640,835, 4,496,689, 4,301,144, 4,670,417,4,791,192 and 4,179,337. In certain embodiments, a derivative bindingagent comprises one or more of monomethoxy-polyethylene glycol, dextran,cellulose, or other carbohydrate based polymers, poly-(N-vinylpyrrolidone)-polyethylene glycol, propylene glycol homopolymers, apolypropylene oxide/ethylene oxide co-polymer, polyoxyethylated polyols(e.g., glycerol) and polyvinyl alcohol, as well as mixtures of suchpolymers. In certain embodiments, one or more water-soluble polymer israndomly attached to one or more side chains. In certain embodiments,PEG can act to improve the therapeutic capacity for a binding agent,such as an antibody. Certain such methods are discussed, for example, inU.S. Pat. No. 6,133,426, which is hereby incorporated by reference forany purpose.

It will be appreciated that an antibody of the present invention mayhave at least one amino acid substitution, deletion, or addition,providing that the antibody retains binding specificity. Therefore,modifications to the antibody structures are encompassed within thescope of the invention. These may include amino acid substitutions,which may be conservative or non-conservative, that do not destroy thehuman TSLP binding capability of an antibody. Conservative amino acidsubstitutions may encompass non-naturally occurring amino acid residues,which are typically incorporated by chemical peptide synthesis ratherthan by synthesis in biological systems. These include peptidomimeticsand other reversed or inverted forms of amino acid moieties. Aconservative amino acid substitution may also involve a substitution ofa native amino acid residue with a normative residue such that there islittle or no effect on the polarity or charge of the amino acid residueat that position.

Non-conservative substitutions may involve the exchange of a member ofone class of amino acids or amino acid mimetics for a member fromanother class with different physical properties (e.g. size, polarity,hydrophobicity, charge). Such substituted residues may be introducedinto regions of the human antibody that are homologous with non-humanantibodies, or into the non-homologous regions of the molecule.

Moreover, one skilled in the art may generate test variants containing asingle amino acid substitution at each desired amino acid residue. Thevariants can then be screened using activity assays known to thoseskilled in the art. Such variants could be used to gather informationabout suitable variants. For example, if one discovered that a change toa particular amino acid residue resulted in destroyed, undesirablyreduced, or unsuitable activity, variants with such a change may beavoided. In other words, based on information gathered from such routineexperiments, one skilled in the art can readily determine the aminoacids where further substitutions should be avoided either alone or incombination with other mutations.

A skilled artisan will be able to determine suitable variants of thepolypeptide as set forth herein using well-known techniques. In certainembodiments, one skilled in the art may identify suitable areas of themolecule that may be changed without destroying activity by targetingregions not believed to be important for activity. In certainembodiments, one can identify residues and portions of the moleculesthat are conserved among similar polypeptides. In certain embodiments,even areas that may be important for biological activity or forstructure may be subject to conservative amino acid substitutionswithout destroying the biological activity or without adverselyaffecting the polypeptide structure.

Additionally, one skilled in the art can review structure-functionstudies identifying residues in similar polypeptides that are importantfor activity or structure. In view of such a comparison, one can predictthe importance of amino acid residues in a protein that correspond toamino acid residues which are important for activity or structure insimilar proteins. One skilled in the art may opt for chemically similaramino acid substitutions for such predicted important amino acidresidues.

One skilled in the art can also analyze the three-dimensional structureand amino acid sequence in relation to that structure in similarpolypeptides. In view of such information, one skilled in the art maypredict the alignment of amino acid residues of an antibody with respectto its three dimensional structure. In certain embodiments, one skilledin the art may choose not to make radical changes to amino acid residuespredicted to be on the surface of the protein, since such residues maybe involved in important interactions with other molecules.

A number of scientific publications have been devoted to the predictionof secondary structure. See Moult J., Curr. Op. in Biotech.,7(4):422-427 (1996), Chou et al., Biochemistry, 13(2):222-245 (1974);Chou et al., Biochemistry, 113(2):211-222 (1974); Chou et al., Adv.Enzymol. Relat. Areas Mol. Biol., 47:45-148 (1978); Chou et al., Ann.Rev. Biochem., 47:251-276 and Chou et al., Biophys. J., 26:367-384(1979). Moreover, computer programs are currently available to assistwith predicting secondary structure. One method of predicting secondarystructure is based upon homology modeling. For example, two polypeptidesor proteins which have a sequence identity of greater than 30%, orsimilarity greater than 40% often have similar structural topologies.The recent growth of the protein structural database (PDB) has providedenhanced predictability of secondary structure, including the potentialnumber of folds within a polypeptide's or protein's structure. See Holmet al., Nucl. Acid. Res., 27(1):244-247 (1999). It has been suggested(Brenner et al., Curr. Op. Struct. Biol., 7(3):369-376 (1997)) thatthere are a limited number of folds in a given polypeptide or proteinand that once a critical number of structures have been resolved,structural prediction will become dramatically more accurate.

Additional methods of predicting secondary structure include “threading”(Jones, D., Curr. Opin. Struct. Biol., 7(3):377-87 (1997); Sippl et al.,Structure, 4(1):15-19 (1996)), “profile analysis” (Bowie et al.,Science, 253:164-170 (1991); Gribskov et al., Meth. Enzym., 183:146-159(1990); Gribskov et al., Proc. Nat. Acad. Sci., 84(13):4355-4358(1987)), and “evolutionary linkage” (See Holm, supra (1999), andBrenner, supra (1997)).

It will be understood by one skilled in the art that some proteins, suchas antibodies, may undergo a variety of posttranslational modifications.The type and extent of these modifications often depends on the hostcell line used to express the protein as well as the culture conditions.Such modifications may include variations in glycosylation, methionineoxidation, diketopiperizine formation, aspartate isomerization andasparagine deamidation. A frequent modification is the loss of acarboxy-terminal basic residue (such as lysine or arginine) due to theaction of carboxypeptidases (as described in Harris, R. J. Journal ofChromatography 705:129-134, 1995).

In certain embodiments, variants of antibodies include glycosylationvariants wherein the number and/or type of glycosylation site has beenaltered compared to the amino acid sequences of a parent polypeptide. Incertain embodiments, variants comprise a greater or a lesser number ofN-linked glycosylation sites than the native protein. Alternatively,substitutions which eliminate this sequence will remove an existingN-linked carbohydrate chain. Also provided is a rearrangement ofN-linked carbohydrate chains wherein one or more N-linked glycosylationsites (typically those that are naturally occurring) are eliminated andone or more new N-linked sites are created. Additional preferredantibody variants include cysteine variants wherein one or more cysteineresidues are deleted from or substituted for another amino acid (e.g.,serine) as compared to the parent amino acid sequence. Cysteine variantsmay be useful when antibodies must be refolded into a biologicallyactive conformation such as after the isolation of insoluble inclusionbodies. Cysteine variants generally have fewer cysteine residues thanthe native protein, and typically have an even number to minimizeinteractions resulting from unpaired cysteines.

Desired amino acid substitutions (whether conservative ornon-conservative) can be determined by those skilled in the art at thetime such substitutions are desired. In certain embodiments, amino acidsubstitutions can be used to identify important residues of antibodiesto human TSLP, or to increase or decrease the affinity of the antibodiesto human TSLP described herein.

According to certain embodiments, preferred amino acid substitutions arethose which: (1) reduce susceptibility to proteolysis, (2) reducesusceptibility to oxidation, (3) alter binding affinity for formingprotein complexes, (4) alter binding affinities, and/or (4) confer ormodify other physiochemical or functional properties on suchpolypeptides. According to certain embodiments, single or multiple aminoacid substitutions (in certain embodiments, conservative amino acidsubstitutions) may be made in the naturally-occurring sequence (incertain embodiments, in the portion of the polypeptide outside thedomain(s) forming intermolecular contacts). In certain embodiments, aconservative amino acid substitution typically may not substantiallychange the structural characteristics of the parent sequence (e.g., areplacement amino acid should not tend to break a helix that occurs inthe parent sequence, or disrupt other types of secondary structure thatcharacterizes the parent sequence). Examples of art-recognizedpolypeptide secondary and tertiary structures are described in Proteins,Structures and Molecular Principles (Creighton, Ed., W. H. Freeman andCompany, New York (1984)); Introduction to Protein Structure (C. Brandenand J. Tooze, eds., Garland Publishing, New York, N.Y. (1991)); andThornton et al. Nature 354:105 (1991), which are each incorporatedherein by reference.

In certain embodiments, antibodies of the invention may be chemicallybonded with polymers, lipids, or other moieties.

In addition, the antigen binding proteins may comprise at least one ofthe CDRs described herein incorporated into a biocompatible frameworkstructure. In one example, the biocompatible framework structurecomprises a polypeptide or portion thereof that is sufficient to form aconformationally stable structural support, or framework, or scaffold,which is able to display one or more sequences of amino acids that bindto an antigen (e.g., CDRs, a variable region, etc.) in a localizedsurface region. Such structures can be a naturally occurring polypeptideor polypeptide “fold” (a structural motif), or can have one or moremodifications, such as additions, deletions or substitutions of aminoacids, relative to a naturally occurring polypeptide or fold. Thesescaffolds can be derived from a polypeptide of any species (or of morethan one species), such as a human, other mammal, other vertebrate,invertebrate, plant, bacteria or virus.

Typically the biocompatible framework structures are based on proteinscaffolds or skeletons other than immunoglobulin domains. For example,those based on fibronectin, ankyrin, lipocalin, neocarzinostain,cytochrome b, CPI zinc finger, PST1, coiled coil, LACI-D1, Z domain andtendamistat domains may be used (See e.g., Nygren and Uhlen, 1997,Current Opinion in Structural Biology, 7, 463-469).

Additionally, in another embodiment, one skilled in the art willrecognize that the antigen binding proteins can include one or more ofheavy chain CDR1, CDR2, CDR3, and/or light chain CDR1, CDR2 and CDR3having one amino acid substitution, provided that the antibody retainsthe binding specificity of the non-substituted CDR. The non-CDR portionof the antibody may be a non-protein molecule, wherein the binding agentcross-blocks the binding of an antibody disclosed herein to human TSLPand/or inhibits TSLP activity. The non-CDR portion of the antibody maybe a non-protein molecule in which the antibody exhibits a similarbinding pattern to human TSLP proteins in a competition binding assay asthat exhibited by at least one of antibodies A1-A27, and/or neutralizesthe activity of TSLP. The non-CDR portion of the antibody may becomposed of amino acids, wherein the antibody is a recombinant bindingprotein or a synthetic peptide, and the recombinant binding proteincross-blocks the binding of an antibody disclosed herein to human TSLPand/or neutralizes TSLP in vitro or in vivo. The non-CDR portion of theantibody may be composed of amino acids, wherein the antibody is arecombinant antibody, and the recombinant antibody exhibits a similarbinding pattern to human TSLP polypeptides in a competition bindingassay as exhibited by at least one of the antibodies A1-A27, and/orneutralizes TSLP activity.

Methods of Making Antigen Binding Proteins, Specifically Antibodies.

An antigen binding protein such as an antibody comprising one or more ofheavy chain CDR1, CDR2, CDR3, and/or light chain CDR1, CDR2 and CDR3 asdescribed above, may be obtained by expression from a host cellcontaining DNA coding for these sequences. A DNA coding for each CDRsequence may be determined on the basis of the amino acid sequence ofthe CDR and synthesized together with any desired antibody variableregion framework and constant region DNA sequences using oligonucleotidesynthesis techniques, site-directed mutagenesis and polymerase chainreaction (PCR) techniques as appropriate. DNA coding for variable regionframeworks and constant regions is widely available to those skilled inthe art from genetic sequences databases such as GenBank®.

Additional embodiments include chimeric antibodies, e.g., humanizedversions of non-human (e.g., murine) monoclonal antibodies. Suchhumanized antibodies may be prepared by known techniques, and offer theadvantage of reduced immunogenicity when the antibodies are administeredto humans. In one embodiment, a humanized monoclonal antibody comprisesthe variable domain of a murine antibody (or all or part of the antigenbinding site thereof) and a constant domain derived from a humanantibody. Alternatively, a humanized antibody fragment may comprise theantigen binding site of a murine monoclonal antibody and a variabledomain fragment (lacking the antigen-binding site) derived from a humanantibody. Procedures for the production of chimeric and furtherengineered monoclonal antibodies include those described in Riechmann etal., 1988, Nature 332:323, Liu et al., 1987, Proc. Nat. Acad. Sci. USA84:3439, Larrick et al., 1989, Bio/Technology 7:934, and Winter et al.,1993, TIPS 14:139. In one embodiment, the chimeric antibody is a CDRgrafted antibody. Techniques for humanizing antibodies are discussed in,e.g., U.S. Pat. Nos. 5,869,619, 5,225,539, 5,821,337, 5,859,205,6,881,557, Padlan et al., 1995, FASEB J. 9:133-39, and Tamura et al.,2000, J. Immunol. 164:1432-41. Addition techniques for producinghumanized antibodies such as those are described in Zhang, W., et al.,Molecular Immunology. 42(12):1445-1451, 2005; Hwang W. et al., Methods.36(1):35-42, 2005; Dall'Acqua W F, et al., Methods 36(1):43-60, 2005;and Clark, M., Immunology Today. 21(8):397-402, 2000).

Procedures have been developed for generating human or partially humanantibodies in non-human animals. For example, mice in which one or moreendogenous immunoglobulin genes have been inactivated by various meanshave been prepared. Human immunoglobulin genes have been introduced intothe mice to replace the inactivated mouse genes. Antibodies produced inthe animal incorporate human immunoglobulin polypeptide chains encodedby the human genetic material introduced into the animal. In oneembodiment, a non-human animal, such as a transgenic mouse, is immunizedwith TSLP protein, for example, such that antibodies directed againstvarious TSLP polypeptides are generated in the animal. Examples ofsuitable immunogens are provided in the Examples below.

Examples of techniques for production and use of transgenic animals forthe production of human or partially human antibodies are described inU.S. Pat. Nos. 5,814,318, 5,569,825, and 5,545,806, Davis et al., 2003,Production of human antibodies from transgenic mice in Lo, ed. AntibodyEngineering Methods and Protocols, Humana Press, NJ: 191-200, Kellermannet al., 2002, Curr Opin Biotechnol. 13:593-97, Russel et al., 2000,Infect Immun. 68:1820-26, Gallo et al., 2000, Eur J Immun. 30:534-40,Davis et al., 1999, Cancer Metastasis Rev. 18:421-25, Green, 1999, JImmunol Methods. 231:11-23, Jakobovits, 1998, Advanced Drug DeliveryReviews 31:33-42, Green et al., 1998, J Exp Med. 188:483-95, JakobovitsA, 1998, Exp. Opin. Invest. Drugs. 7:607-14, Tsuda et al., 1997,Genomics. 42:413-21, Mendez et al., 1997, Nat. Genet. 15:146-56,Jakobovits, 1994, Curr Biol. 4:761-63, Arbones et al., 1994, Immunity.1:247-60, Green et al., 1994, Nat. Genet. 7:13-21, Jakobovits et al.,1993, Nature. 362:255-58, Jakobovits et al., 1993, Proc Natl Acad SciUSA. 90:2551-55. Chen, J., M. Trounstine, F. W. Alt, F. Young, C.Kurahara, J. Loring, D. Huszar. “Immunoglobulin gene rearrangement inB-cell deficient mice generated by targeted deletion of the JH locus.”International Immunology 5 (1993): 647-656, Choi et al., 1993, NatureGenetics 4: 117-23, Fishwild et al., 1996, Nature Biotechnology 14:845-51, Harding et al., 1995, Annals of the New York Academy ofSciences, Lonberg et al., 1994, Nature 368: 856-59, Lonberg, 1994,Transgenic Approaches to Human Monoclonal Antibodies in Handbook ofExperimental Pharmacology 113: 49-101, Lonberg et al., 1995, InternalReview of Immunology 13: 65-93, Neuberger, 1996, Nature Biotechnology14: 826, Taylor et al., 1992, Nucleic Acids Research 20: 6287-95, Tayloret al., 1994, International Immunology 6: 579-91, Tomizuka et al., 1997,Nature Genetics 16: 133-43, Tomizuka et al., 2000, Proceedings of theNational Academy of Sciences USA 97: 722-27, Tuaillon et al., 1993,Proceedings of the National Academy of Sciences USA 90: 3720-24, andTuaillon et al., 1994, Journal of Immunology 152: 2912-20.

In another aspect, the present invention provides monoclonal antibodiesthat bind to human TSLP. Monoclonal antibodies may be produced using anytechnique known in the art, e.g., by immortalizing spleen cellsharvested from the transgenic animal after completion of theimmunization schedule. The spleen cells can be immortalized using anytechnique known in the art, e.g., by fusing them with myeloma cells toproduce hybridomas. Myeloma cells for use in hybridoma-producing fusionprocedures preferably are non-antibody-producing, have high fusionefficiency, and enzyme deficiencies that render them incapable ofgrowing in certain selective media which support the growth of only thedesired fused cells (hybridomas). Examples of suitable cell lines foruse in mouse fusions include Sp-20, P3-X63/Ag8, P3-X63-Ag8.653, NS1/1.Ag4 1, Sp210-Ag14, FO, NSO/U, MPC-11, MPC11-X45-GTG 1.7 and S194/5XX0 Bul;examples of cell lines used in rat fusions include R210.RCY3, Y3-Ag1.2.3, IR983F and 4B210. Other cell lines useful for cell fusions areU-266, GM 1500-GRG2, LICR-LON-HMy2 and UC729-6.

In one embodiment, a hybridoma cell line is produced by immunizing ananimal (e.g., a transgenic animal having human immunoglobulin sequences)with a TSLP immunogen; harvesting spleen cells from the immunizedanimal; fusing the harvested spleen cells to a myeloma cell line,thereby generating hybridoma cells; establishing hybridoma cell linesfrom the hybridoma cells, and identifying a hybridoma cell line thatproduces an antibody that binds a TSLP polypeptide. Such hybridoma celllines, and TSLP monoclonal antibodies produced by them, are encompassedby the present invention.

Monoclonal antibodies secreted by a hybridoma cell line can be purifiedusing any technique known in the art. Hybridomas or mAbs may be furtherscreened to identify mAbs with particular properties, such as blocking aTSLP activity such as osteoprotegerin (OPG) production from primaryhuman dendritic cells. Examples of such assays are provided in theexamples below.

Molecular evolution of the complementarity determining regions (CDRs) inthe center of the antibody binding site also has been used to isolateantibodies with increased affinity, for example, as described by Schieret al., 1996, J. Mol. Biol. 263:551. Accordingly, such techniques areuseful in preparing antibodies to human TSLP.

Antigen binding proteins directed against human TSLP can be used, forexample, in assays to detect the presence of TSLP either in vitro or invivo.

Although human, partially human, or humanized antibodies will besuitable for many applications, particularly those involvingadministration of the antibody to a human subject, other types ofantigen binding proteins will be suitable for certain applications. Thenon-human antibodies of the invention can be, for example, derived fromany antibody-producing animal, such as mouse, rat, rabbit, goat, donkey,or non-human primate (such as monkey (e.g., cynomologus or rhesusmonkey) or ape (e.g., chimpanzee)). Non-human antibodies of theinvention can be used, for example, in in vitro and cell-culture basedapplications, or any other application where an immune response to theantibody of the invention does not occur, is insignificant, can beprevented, is not a concern, or is desired. In one embodiment, anon-human antibody of the invention is administered to a non-humansubject. In another embodiment, the non-human antibody does not elicitan immune response in the non-human subject. In another embodiment, thenon-human antibody is from the same species as the non-human subject,e.g., a mouse antibody of the invention is administered to a mouse. Anantibody from a particular species can be made by, for example,immunizing an animal of that species with the desired immunogen or usingan artificial system for generating antibodies of that species (e.g., abacterial or phage display-based system for generating antibodies of aparticular species), or by converting an antibody from one species intoan antibody from another species by replacing, e.g., the constant regionof the antibody with a constant region from the other species, or byreplacing one or more amino acid residues of the antibody so that itmore closely resembles the sequence of an antibody from the otherspecies. In one embodiment, the antibody is a chimeric antibodycomprising amino acid sequences derived from antibodies from two or moredifferent species.

Antigen binding proteins may be prepared by any of a number ofconventional techniques. For example, they may be purified from cellsthat naturally express them (e.g., an antibody can be purified from ahybridoma that produces it), or produced in recombinant expressionsystems, using any technique known in the art. See, for example,Monoclonal Antibodies, Hybridomas: A New Dimension in BiologicalAnalyses, Kennet et al. (eds.), Plenum Press, New York (1980); andAntibodies: A Laboratory Manual, Harlow and Land (eds.), Cold SpringHarbor Laboratory Press, Cold Spring Harbor, N.Y., (1988).

Any expression system known in the art can be used to make therecombinant polypeptides of the invention. In general, host cells aretransformed with a recombinant expression vector that comprises DNAencoding a desired polypeptide. Among the host cells that may beemployed are prokaryotes, yeast or higher eukaryotic cells. Prokaryotesinclude gram negative or gram positive organisms, for example E. coli orbacilli. Higher eukaryotic cells include insect cells and establishedcell lines of mammalian origin. Examples of suitable mammalian host celllines include the COS-7 line of monkey kidney cells (ATCC CRL 1651)(Gluzman et al., 1981, Cell 23:175), L cells, 293 cells, C127 cells, 3T3cells (ATCC CCL 163), Chinese hamster ovary (CHO) cells, HeLa cells, BHK(ATCC CRL 10) cell lines, and the CVI/EBNA cell line derived from theAfrican green monkey kidney cell line CV1 (ATCC CCL 70) as described byMcMahan et al., 1991, EMBO J. 10: 2821. Appropriate cloning andexpression vectors for use with bacterial, fungal, yeast, and mammaliancellular hosts are described by Pouwels et al. (Cloning Vectors: ALaboratory Manual, Elsevier, N.Y., 1985).

The transformed cells can be cultured under conditions that promoteexpression of the polypeptide, and the polypeptide recovered byconventional protein purification procedures. One such purificationprocedure is described in the Examples below. Polypeptides contemplatedfor use herein include substantially homogeneous recombinant mammaliananti-TSLP antibody polypeptides substantially free of contaminatingendogenous materials.

Antigen binding proteins may be prepared, and screened for desiredproperties, by any of a number of known techniques. Certain of thetechniques involve isolating a nucleic acid encoding a polypeptide chain(or portion thereof) of an antigen binding protein of interest (e.g., anTSLP antibody), and manipulating the nucleic acid through recombinantDNA technology. The nucleic acid may be fused to another nucleic acid ofinterest, or altered (e.g., by mutagenesis or other conventionaltechniques) to add, delete, or substitute one or more amino acidresidues, for example.

Single chain antibodies may be formed by linking heavy and light chainvariable domain (Fv region) fragments via an amino acid bridge (shortpeptide linker), resulting in a single polypeptide chain. Suchsingle-chain Fvs (scFvs) have been prepared by fusing DNA encoding apeptide linker between DNAs encoding the two variable domainpolypeptides (V_(L) and V_(H)). The resulting polypeptides can fold backon themselves to form antigen-binding monomers, or they can formmultimers (e.g., dimers, trimers, or tetramers), depending on the lengthof a flexible linker between the two variable domains (Kortt et al.,1997, Prot. Eng. 10:423; Kortt et al., 2001, Biomol. Eng. 18:95-108). Bycombining different V_(L) and V_(H)-comprising polypeptides, one canform multimeric scFvs that bind to different epitopes (Kriangkum et al.,2001, Biomol. Eng. 18:31-40). Techniques developed for the production ofsingle chain antibodies include those described in U.S. Pat. No.4,946,778; Bird, 1988, Science 242:423; Huston et al., 1988, Proc. Natl.Acad. Sci. USA 85:5879; Ward et al., 1989, Nature 334:544, de Graaf etal., 2002, Methods Mol. Biol. 178:379-87. Single chain antibodiesderived from antibodies provided herein include, but are not limited to,scFvs comprising the variable domain combinations L1H1, L2H2, L3H3,L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13H13,L14H14, L15H15, L16H16, L17H17, L18H18, L19H19, L20H20, L21H21, L22H22,L23H23, L24H24, L25H25, L26H26, and L27H27 are encompassed by thepresent invention.

Once synthesized, the DNA encoding an antibody of the invention orfragment thereof may be propagated and expressed according to any of avariety of well-known procedures for nucleic acid excision, ligation,transformation, and transfection using any number of known expressionvectors. Thus, in certain embodiments expression of an antibody fragmentmay be preferred in a prokaryotic host, such as Escherichia coli (see,e.g., Pluckthun et al., 1989 Methods Enzymol. 178:497-515). In certainother embodiments, expression of the antibody or a fragment thereof maybe preferred in a eukaryotic host cell, including yeast (e.g.,Saccharomyces cerevisiae, Schizosaccharomyces pombe, and Pichiapastoris), animal cells (including mammalian cells) or plant cells.Examples of suitable animal cells include, but are not limited to,myeloma (such as a mouse NSO line), COS, CHO, or hybridoma cells.Examples of plant cells include tobacco, corn, soybean, and rice cells.One or more replicable expression vectors containing DNA encoding anantibody variable and/or constant region may be prepared and used totransform an appropriate cell line, for example, a non-producing myelomacell line, such as a mouse NSO line or a bacteria, such as E. coli, inwhich production of the antibody will occur. In order to obtainefficient transcription and translation, the DNA sequence in each vectorshould include appropriate regulatory sequences, particularly a promoterand leader sequence operatively linked to the variable domain sequence.Particular methods for producing antibodies in this way are generallywell-known and routinely used. For example, basic molecular biologyprocedures are described by Maniatis et al. (Molecular Cloning, ALaboratory Manual, 2nd ed., Cold Spring Harbor Laboratory, New York,1989; see also Maniatis et al, 3rd ed., Cold Spring Harbor Laboratory,New York, (2001)). DNA sequencing can be performed as described inSanger et al. (PNAS 74:5463, (1977)) and the Amersham International plcsequencing handbook, and site directed mutagenesis can be carried outaccording to methods known in the art (Kramer et al., Nucleic Acids Res.12:9441, (1984); Kunkel Proc. Natl. Acad. Sci. USA 82:488-92 (1985);Kunkel et al., Methods in Enzymol. 154:367-82 (1987); the AnglianBiotechnology Ltd. handbook). Additionally, numerous publicationsdescribe techniques suitable for the preparation of antibodies bymanipulation of DNA, creation of expression vectors, and transformationand culture of appropriate cells (Mountain A and Adair, J R inBiotechnology and Genetic Engineering Reviews (ed. Tombs, M P, 10,Chapter 1, 1992, Intercept, Andover, UK); “Current Protocols inMolecular Biology”, 1999, F. M. Ausubel (ed.), Wiley Interscience, NewYork).

Where it is desired to improve the affinity of antibodies according tothe invention containing one or more of the above-mentioned CDRs can beobtained by a number of affinity maturation protocols includingmaintaining the CDRs (Yang et al., J. Mol. Biol., 254, 392-403, 1995),chain shuffling (Marks et al., Bio/Technology, 10, 779-783, 1992), useof mutation strains of E. coli. (Low et al., J. Mol. Biol., 250,350-368, 1996), DNA shuffling (Patten et al., Curr. Opin. Biotechnol.,8, 724-733, 1997), phage display (Thompson et al., J. Mol. Biol., 256,7-88, 1996) and PCR (Crameri, et al., Nature, 391, 288-291, 1998). Allof these methods of affinity maturation are discussed by Vaughan et al.(Nature Biotechnology, 16, 535-539, 1998).

Other antibodies according to the invention may be obtained byconventional immunization and cell fusion procedures as described hereinand known in the art. Monoclonal antibodies of the invention may begenerated using a variety of known techniques. In general, monoclonalantibodies that bind to specific antigens may be obtained by methodsknown to those skilled in the art (see, for example, Kohler et al.,Nature 256:495, 1975; Coligan et al. (eds.), Current Protocols inImmunology, 1:2.5.12.6.7 (John Wiley & Sons 1991); U.S. Pat. Nos. RE32,011, 4,902,614, 4,543,439, and 4,411,993; Monoclonal Antibodies,Hybridomas: A New Dimension in Biological Analyses, Plenum Press,Kennett, McKearn, and Bechtol (eds.) (1980); and Antibodies: ALaboratory Manual, Harlow and Lane (eds.), Cold Spring Harbor LaboratoryPress (1988); Picksley et al., “Production of monoclonal antibodiesagainst proteins expressed in E. coli,” in DNA Cloning 2: ExpressionSystems, 2nd Edition, Glover et al. (eds.), page 93 (Oxford UniversityPress 1995)). Antibody fragments may be derived therefrom using anysuitable standard technique such as proteolytic digestion, oroptionally, by proteolytic digestion (for example, using papain orpepsin) followed by mild reduction of disulfide bonds and alkylation.Alternatively, such fragments may also be generated by recombinantgenetic engineering techniques as described herein.

Monoclonal antibodies can be obtained by injecting an animal, forexample, a rat, hamster, a rabbit, or preferably a mouse, including forexample a transgenic or a knock-out, as known in the art, with animmunogen comprising human TSLP of SEQ ID NO: 2, other TSLP polypeptidesequences as described herein, or a fragment thereof, according tomethods known in the art and described herein. The presence of specificantibody production may be monitored after the initial injection and/orafter a booster injection by obtaining a serum sample and detecting thepresence of an antibody that binds to human TSLP or fragment thereofusing any one of several immunodetection methods known in the art anddescribed herein. From animals producing the desired antibodies,lymphoid cells, most commonly cells from the spleen or lymph node, areremoved to obtain B-lymphocytes. The B lymphocytes are then fused with adrug-sensitized myeloma cell fusion partner, preferably one that issyngeneic with the immunized animal and that optionally has otherdesirable properties (e.g., inability to express endogenous Ig geneproducts, e.g., P3×63-Ag 8.653 (ATCC No. CRL 1580); NSO, SP20) toproduce hybridomas, which are immortal eukaryotic cell lines.

The lymphoid (e.g., spleen) cells and the myeloma cells may be combinedfor a few minutes with a membrane fusion-promoting agent, such aspolyethylene glycol or a nonionic detergent, and then plated at lowdensity on a selective medium that supports the growth of hybridomacells but not unfused myeloma cells. A preferred selection media is HAT(hypoxanthine, aminopterin, thymidine). After a sufficient time, usuallyabout one to two weeks, colonies of cells are observed. Single coloniesare isolated, and antibodies produced by the cells may be tested forbinding activity to human TSLP using any one of a variety ofimmunoassays known in the art and described herein. The hybridomas arecloned (e.g., by limited dilution cloning or by soft agar plaqueisolation) and positive clones that produce an antibody specific tohuman TSLP are selected and cultured. The monoclonal antibodies from thehybridoma cultures may be isolated from the supernatants of hybridomacultures.

An alternative method for production of a murine monoclonal antibody isto inject the hybridoma cells into the peritoneal cavity of a syngeneicmouse, for example, a mouse that has been treated (e.g.,pristane-primed) to promote formation of ascites fluid containing themonoclonal antibody. Monoclonal antibodies can be isolated and purifiedby a variety of well-established techniques. Such isolation techniquesinclude affinity chromatography with Protein-A Sepharose, size-exclusionchromatography, and ion-exchange chromatography (see, for example,Coligan at pages 2.7.1-2.7.12 and pages 2.9.1-2.9.3; Baines et al.,“Purification of Immunoglobulin G (IgG),” in Methods in MolecularBiology, Vol. 10, pages 79-104 (The Humana Press, Inc. 1992)).Monoclonal antibodies may be purified by affinity chromatography usingan appropriate ligand selected based on particular properties of theantibody (e.g., heavy or light chain isotype, binding specificity,etc.). Examples of a suitable ligand, immobilized on a solid support,include Protein A, Protein G, an anticonstant region (light chain orheavy chain) antibody, an anti-idiotype antibody, and TSLP, or fragmentor variant thereof.

An antibody of the present invention may also be a fully humanmonoclonal antibody. Fully human monoclonal antibodies may be generatedby any number of techniques as those previsously described above. Suchmethods further include, but are not limited to, Epstein Barr Virus(EBV) transformation of human peripheral blood cells (e.g., containing Blymphocytes), in vitro immunization of human B-cells, fusion of spleencells from immunized transgenic mice carrying inserted humanimmunoglobulin genes, isolation from human immunoglobulin V region phagelibraries, or other procedures as known in the art and based on thedisclosure herein. For example, fully human monoclonal antibodies may beobtained from transgenic mice that have been engineered to producespecific human antibodies in response to antigenic challenge. Methodsfor obtaining fully human antibodies from transgenic mice are described,for example, by Green et al., Nature Genet. 7:13, 1994; Lonberg et al.,Nature 368:856, 1994; Taylor et al., Int. Immun. 6:579, 1994; U.S. Pat.No. 5,877,397; Bruggemann et al., 1997 Curr. Opin. Biotechnol. 8:455-58;Jakobovits et al., 1995 Ann. N.Y. Acad. Sci. 764:525-35. In thistechnique, elements of the human heavy and light chain locus areintroduced into strains of mice derived from embryonic stem cell linesthat contain targeted disruptions of the endogenous heavy chain andlight chain loci (see also Bruggemann et al., Curr. Opin. Biotechnol.8:455-58 (1997)). For example, human immunoglobulin transgenes may bemini-gene constructs, or transloci on yeast artificial chromosomes,which undergo B-cell-specific DNA rearrangement and hypermutation in themouse lymphoid tissue. Fully human monoclonal antibodies may be obtainedby immunizing the transgenic mice, which may then produce humanantibodies specific for human TSLP. Lymphoid cells of the immunizedtransgenic mice can be used to produce human antibody-secretinghybridomas according to the methods described herein. Polyclonal seracontaining fully human antibodies may also be obtained from the blood ofthe immunized animals.

One exemplary method for generating human antibodies of the inventionincludes immortalizing human peripheral blood cells by EBVtransformation, as described, for example, in U.S. Pat. No. 4,464,456.Such an immortalized B-cell line (or lymphoblastoid cell line) producinga monoclonal antibody that specifically binds to human TSLP can beidentified by immunodetection methods as provided herein, for example,an ELISA, and then isolated by standard cloning techniques. Thestability of the lymphoblastoid cell line producing an anti-TSLPantibody may be improved by fusing the transformed cell line with amurine myeloma to produce a mouse-human hybrid cell line according tomethods known in the art (see, e.g., Glasky et al., Hybridoma 8:377-89(1989)). Still another method to generate human monoclonal antibodies isin vitro immunization, which includes priming human splenic B-cells withhuman TSLP followed by fusion of primed B-cells with a heterohybridfusion partner. See, e.g., Boerner et al., 1991 J. Immunol. 147:86-95.

In certain embodiments, a B-cell that is producing an anti-human TSLPantibody is selected and the light chain and heavy chain variableregions are cloned from the B-cell according to molecular biologytechniques known in the art (WO 92/02551; U.S. Pat. No. 5,627,052;Babcook et al., Proc. Natl. Acad. Sci. USA 93:7843-48 (1996)) anddescribed herein. B-cells from an immunized animal may be isolated fromthe spleen, lymph node, or peripheral blood sample by selecting a cellthat is producing an antibody that specifically binds to TSLP. B-cellsmay also be isolated from humans, for example, from a peripheral bloodsample. Methods for detecting single B-cells that are producing anantibody with the desired specificity are well known in the art, forexample, by plaque formation, fluorescence-activated cell sorting, invitro stimulation followed by detection of specific antibody, and thelike. Methods for selection of specific antibody-producing B-cellsinclude, for example, preparing a single cell suspension of B-cells insoft agar that contains human TSLP. Binding of the specific antibodyproduced by the B-cell to the antigen results in the formation of acomplex, which may be visible as an immunoprecipitate. After the B-cellsproducing the desired antibody are selected, the specific antibody genesmay be cloned by isolating and amplifying DNA or mRNA according tomethods known in the art and described herein.

An additional method for obtaining antibodies of the invention is byphage display. See, e.g., Winter et al., 1994 Annu. Rev. Immunol.12:433-55; Burton et al., 1994 Adv. Immunol. 57:191-280. Human or murineimmunoglobulin variable region gene combinatorial libraries may becreated in phage vectors that can be screened to select Ig fragments(Fab, Fv, sFv, or multimers thereof) that bind specifically to TSLP orvariant or fragment thereof. See, e.g., U.S. Pat. No. 5,223,409; Huse etal., 1989 Science 246:1275-81; Sastry et al., Proc. Natl. Acad. Sci. USA86:5728-32 (1989); Alting-Mees et al., Strategies in Molecular Biology3:1-9 (1990); Kang et al., 1991 Proc. Natl. Acad. Sci. USA 88:4363-66;Hoogenboom et al., 1992 J. Molec. Biol. 227:381-388; Schlebusch et al.,1997 Hybridoma 16:47-52 and references cited therein. For example, alibrary containing a plurality of polynucleotide sequences encoding Igvariable region fragments may be inserted into the genome of afilamentous bacteriophage, such as M13 or a variant thereof, in framewith the sequence encoding a phage coat protein. A fusion protein may bea fusion of the coat protein with the light chain variable region domainand/or with the heavy chain variable region domain. According to certainembodiments, immunoglobulin Fab fragments may also be displayed on aphage particle (see, e.g., U.S. Pat. No. 5,698,426).

Heavy and light chain immunoglobulin cDNA expression libraries may alsobe prepared in lambda phage, for example, using λImmunoZap™(H) andλImmunoZap™(L) vectors (Stratagene, La Jolla, Calif.). Briefly, mRNA isisolated from a B-cell population, and used to create heavy and lightchain immunoglobulin cDNA expression libraries in the λImmunoZap(H) andλImmunoZap(L) vectors. These vectors may be screened individually orco-expressed to form Fab fragments or antibodies (see Huse et al.,supra; see also Sastry et al., supra). Positive plaques may subsequentlybe converted to a non-lytic plasmid that allows high level expression ofmonoclonal antibody fragments from E. coli.

In one embodiment, in a hybridoma the variable regions of a geneexpressing a monoclonal antibody of interest are amplified usingnucleotide primers. These primers may be synthesized by one of ordinaryskill in the art, or may be purchased from commercially availablesources. (See, e.g., Stratagene (La Jolla, Calif.), which sells primersfor mouse and human variable regions including, among others, primersfor V_(Ha), V_(Hb), V_(Hc), V_(Hd), C_(HI), V_(L) and C_(L) regions.)These primers may be used to amplify heavy or light chain variableregions, which may then be inserted into vectors such as ImmunoZAP™ orImmunoZAP™L (Stratagene), respectively. These vectors may then beintroduced into E. coli, yeast, or mammalian-based systems forexpression. Large amounts of a single-chain protein containing a fusionof the V_(H) and V_(L) domains may be produced using these methods (seeBird et al., Science 242:423-426, 1988).

Once cells producing antibodies according to the invention have beenobtained using any of the above-described immunization and othertechniques, the specific antibody genes may be cloned by isolating andamplifying DNA or mRNA therefrom according to standard procedures asdescribed herein. The antibodies produced therefrom may be sequenced andthe CDRs identified and the DNA coding for the CDRs may be manipulatedas described previously to generate other antibodies according to theinvention.

Antigen binding proteins of the present invention preferably modulateTSLP activity in one of the cell-based assay described herein and/or thein vivo assay described herein and/or cross-block the binding of one ofthe antibodies described in this application and/or are cross-blockedfrom binding TSLP by one of the antibodies described in thisapplication. Particularly useful are antigen binding proteins thatcross-compete with an exemplary antibody described herein, i.e.,cross-block the binding of one of the exemplary antibodies described inthis application and are cross-blocked from binding TSLP by one of theexemplary antibodies. Accordingly such binding agents can be identifiedusing the assays described herein.

In certain embodiments, antibodies are generated by first identifyingantibodies that bind to TSLP and/or neutralize in the cell-based assaysdescribed herein and/or cross-block the antibodies described in thisapplication and/or are cross-blocked from binding TSLP by one of theantibodies described in this application. The CDR regions from theseantibodies are then used to insert into appropriate biocompatibleframeworks to generate antigen binding proteins. The non-CDR portion ofthe binding agent may be composed of amino acids, or may be anon-protein molecule. The assays described herein allow thecharacterization of binding agents. Preferably the binding agents of thepresent invention are antibodies as defined herein.

Antigen binding proteins of the present invention include those thatbind to the same epitope as an exemplary antibody described herein. Asdiscussed in Example 9, epitopes may be structural or functional.Structural epitopes may be thought of as the patch of the target whichis covered by the antibody. Functional epitopes are a subset of thestructural epitopes and comprise those residues which directlycontribute to the affinity of the interaction (e.g. hydrogen bonds,ionic interactions). One method of determining the epitope of anantibody is by using scanning mutations in the target molecule andmeasuring the effect of the mutation on binding. Given thethree-dimensional structure of the antibody binding region, mutations inthe epitope can decrease or increase the binding affinity of theantibody for the mutated target.

Antigen binding proteins may be defined by their epitopes. As seen inTable 6, although the antibodies may all bind to TSLP, they are affecteddifferently by the mutation of certain residues in TSLP an indicationthat their respective epitopes do not completely overlap. Preferredantigen binding proteins include those that share at least a portion ofthe structural epitope of a reference antibody described herein.

For example, a preferred antigen binding protein is one that shares atleast a portion of the same structural epitope as A2. This is evidencedby an increase in binding affinity as compared to for wild-type TSLPwhen TSLP has mutation K67E, K97E, K98E, R100E, K101E, or K103E. Thismay also be evidenced by a decrease in binding affinity as compared tofor wild-type TSLP when TSLP has mutation K21E, T25R, S28R, S64R, orK73E. Although the antigen binding protein and A2 may be affectedsimilarly by some mutations and not others, the more identity there isbetween the antigen binding protein and A2 on the effect of mutations incertain residues of TSLP, the more the antigen binding protein andreference antibody share a structural epitope.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A4. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K97E, K98E, R100E, K101E, or K103E. This may also beevidenced by a decrease in binding affinity as compared to for wild-typeTSLP when TSLP has mutation K10E, A14R, K21E, D22R, K73E, K75E, or A76R.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A5. This is evidenced by adecrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K12E, D22R, S40R, R122E, N124E, R125E, or K129E.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A6. This is evidenced by adecrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation S40R, S42R, H46R, R122E, or K129E.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A7. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K101E. This may also be evidenced by a decrease in bindingaffinity as compared to for wild-type TSLP when TSLP has mutation D2R,T4R, D7R, S42R, H46R, T49R, E50R, Q112R, R122E, R125E, or K129E.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A10. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K97E, K98E, R100E, K101E, or K103E. This may also beevidenced by a decrease in binding affinity as compared to for wild-typeTSLP when TSLP has mutation N5R, S17R, Ti 8R, K21E, D22R, T25R, T33R,H46R, A63R, S64R, A66R, E68R, K73E, K75E, A76R, A92R, T93R, Q94R, orA95R.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A21. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K97E, K98E, R100E, K101E, or K103E. This may also beevidenced by a decrease in binding affinity as compared to for wild-typeTSLP when TSLP has mutation K21E, K21R, D22R, T25R, T33R, S64R, K73E,K75E, E111R, or S114R.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A23. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K67E, K97E, K98E, R100E, K101E, or K103E. This may also beevidenced by a decrease in binding affinity as compared to for wild-typeTSLP when TSLP has mutation E9R, K10E, K12E, A13R, S17R, S20R, K21E,K21R, K73E, K75E, N124E, or R125E.

Another preferred antigen binding protein is one that shares at least aportion of the same structural epitope as A26. This is evidenced by anincrease in binding affinity as compared to for wild-type TSLP when TSLPhas mutation K97E, K98E, R100E, K101E, or K103E. This may also beevidenced by a decrease in binding affinity as compared to for wild-typeTSLP when TSLP has mutation A14R, K21E, D22R, A63R, S64R, K67E, K73E,A76R, A92R, or A95R.

Comparing the mutations that affect binding amongst the antibody, itsuggests that certain residues of TSLP tend to be part of the antibodiesability to bind TSLP and block TSLP activity. Such residues include K21,D22, K73, and K129. Thus, preferred antigen binding protein includethose that have a higher affinity for wild-type TSLP than for a TSLPcomprising mutation K21E, those that have a higher affinity forwild-type TSLP than for a TSLP comprising mutation D21R, those that havea higher affinity for wild-type TSLP than for a TSLP comprising mutationK73E, and those that have a higher affinity for wild-type TSLP than fora TSLP comprising mutation K129E.

Furthermore, many of the exemplary antigen binding proteins describedherein share the attribute that the affinity for TSLP increases when thebasic patch of amino acids at positions 97-103 are changed to acidicamino acids.

Nucleic Acids

In one aspect, the present invention provides isolated nucleic acidmolecules. The nucleic acids comprise, for example, polynucleotides thatencode all or part of an antigen binding protein, for example, one orboth chains of an antibody of the invention, or a fragment, derivative,mutein, or variant thereof, polynucleotides sufficient for use ashybridization probes, PCR primers or sequencing primers for identifying,analyzing, mutating or amplifying a polynucleotide encoding apolypeptide, anti-sense nucleic acids for inhibiting expression of apolynucleotide, and complementary sequences of the foregoing. Thenucleic acids can be any length. They can be, for example, 5, 10, 15,20, 25, 30, 35, 40, 45, 50, 75, 100, 125, 150, 175, 200, 250, 300, 350,400, 450, 500, 750, 1,000, 1,500, 3,000, 5,000 or more nucleotides inlength, and/or can comprise one or more additional sequences, forexample, regulatory sequences, and/or be part of a larger nucleic acid,for example, a vector. The nucleic acids can be single-stranded ordouble-stranded and can comprise RNA and/or DNA nucleotides, andartificial variants thereof (e.g., peptide nucleic acids).

Nucleic acids encoding antibody polypeptides (e.g., heavy or lightchain, variable domain only, or full length) may be isolated fromB-cells of mice that have been immunized with a TSLP antigen. Thenucleic acid may be isolated by conventional procedures such aspolymerase chain reaction (PCR).

Nucleic acid sequences encoding the variable regions of the heavy andlight chain variable regions are shown above. The skilled artisan willappreciate that, due to the degeneracy of the genetic code, each of thepolypeptide sequences disclosed herein is encoded by a large number ofother nucleic acid sequences. The present invention provides eachdegenerate nucleotide sequence encoding each antigen binding protein ofthe invention.

The invention further provides nucleic acids that hybridize to othernucleic acids (e.g., nucleic acids comprising a nucleotide sequence ofany of A1-A27) under particular hybridization conditions. Methods forhybridizing nucleic acids are well-known in the art. See, e.g., CurrentProtocols in Molecular Biology, John Wiley & Sons, N.Y. (1989),6.3.1-6.3.6. As defined herein, a moderately stringent hybridizationcondition uses a prewashing solution containing 5× sodiumchloride/sodium citrate (SSC), 0.5% SDS, 1.0 mM EDTA (pH 8.0),hybridization buffer of about 50% formamide, 6×SSC, and a hybridizationtemperature of 55° C. (or other similar hybridization solutions, such asone containing about 50% formamide, with a hybridization temperature of42° C.), and washing conditions of 60° C., in 0.5×SSC, 0.1% SDS. Astringent hybridization condition hybridizes in 6×SSC at 45° C.,followed by one or more washes in 0.1×SSC, 0.2% SDS at 68° C.Furthermore, one of skill in the art can manipulate the hybridizationand/or washing conditions to increase or decrease the stringency ofhybridization such that nucleic acids comprising nucleotide sequencesthat are at least 65, 70, 75, 80, 85, 90, 95, 98 or 99% identical toeach other typically remain hybridized to each other. The basicparameters affecting the choice of hybridization conditions and guidancefor devising suitable conditions are set forth by, for example,Sambrook, Fritsch, and Maniatis (1989, Molecular Cloning: A LaboratoryManual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.,chapters 9 and 11; and Current Protocols in Molecular Biology, 1995,Ausubel et al., eds., John Wiley & Sons, Inc., sections 2.10 and6.3-6.4), and can be readily determined by those having ordinary skillin the art based on, for example, the length and/or base composition ofthe DNA.

Changes can be introduced by mutation into a nucleic acid, therebyleading to changes in the amino acid sequence of a polypeptide (e.g., anantigen binding protein) that it encodes. Mutations can be introducedusing any technique known in the art. In one embodiment, one or moreparticular amino acid residues are changed using, for example, asite-directed mutagenesis protocol. In another embodiment, one or morerandomly selected residues is changed using, for example, a randommutagenesis protocol. However it is made, a mutant polypeptide can beexpressed and screened for a desired property.

Mutations can be introduced into a nucleic acid without significantlyaltering the biological activity of a polypeptide that it encodes. Forexample, one can make nucleotide substitutions leading to amino acidsubstitutions at non-essential amino acid residues. In one embodiment, anucleotide sequence provided herein for A1-A27, or a desired fragment,variant, or derivative thereof, is mutated such that it encodes an aminoacid sequence comprising one or more deletions or substitutions of aminoacid residues that are shown herein for A1-A27 to be residues where twoor more sequences differ. In another embodiment, the mutagenesis insertsan amino acid adjacent to one or more amino acid residues shown hereinfor A1-A27 to be residues where two or more sequences differ.Alternatively, one or more mutations can be introduced into a nucleicacid that selectively change the biological activity. (e.g., binding toTSLP) of a polypeptide that it encodes. For example, the mutation canquantitatively or qualitatively change the biological activity. Examplesof quantitative changes include increasing, reducing or eliminating theactivity. Examples of qualitative changes include changing the antigenspecificity of an antigen binding protein.

In another aspect, the present invention provides nucleic acid moleculesthat are suitable for use as primers or hybridization probes for thedetection of nucleic acid sequences of the invention. A nucleic acidmolecule of the invention can comprise only a portion of a nucleic acidsequence encoding a full-length polypeptide of the invention, forexample, a fragment that can be used as a probe or primer or a fragmentencoding an active portion (e.g., a TSLP binding portion) of apolypeptide of the invention.

Probes based on the sequence of a nucleic acid of the invention can beused to detect the nucleic acid or similar nucleic acids, for example,transcripts encoding a polypeptide of the invention. The probe cancomprise a label group, e.g., a radioisotope, a fluorescent compound, anenzyme, or an enzyme co-factor. Such probes can be used to identify acell that expresses the polypeptide.

In another aspect, the present invention provides vectors comprising anucleic acid encoding a polypeptide of the invention or a portionthereof. Examples of vectors include, but are not limited to, plasmids,viral vectors, non-episomal mammalian vectors and expression vectors,for example, recombinant expression vectors.

The recombinant expression vectors of the invention can comprise anucleic acid of the invention in a form suitable for expression of thenucleic acid in a host cell. The recombinant expression vectors includeone or more regulatory sequences, selected on the basis of the hostcells to be used for expression, which is operably linked to the nucleicacid sequence to be expressed. Regulatory sequences include those thatdirect constitutive expression of a nucleotide sequence in many types ofhost cells (e.g., SV40 early gene enhancer, Rous sarcoma virus promoterand cytomegalovirus promoter), those that direct expression of thenucleotide sequence only in certain host cells (e.g., tissue-specificregulatory sequences, see Voss et al., 1986, Trends Biochem. Sci.11:287, Maniatis et al., 1987, Science 236:1237, incorporated byreference herein in their entireties), and those that direct inducibleexpression of a nucleotide sequence in response to particular treatmentor condition (e.g., the metallothionin promoter in mammalian cells andthe tet-responsive and/or streptomycin responsive promoter in bothprokaryotic and eukaryotic systems (see id.). It will be appreciated bythose skilled in the art that the design of the expression vector candepend on such factors as the choice of the host cell to be transformed,the level of expression of protein desired, etc. The expression vectorsof the invention can be introduced into host cells to thereby produceproteins or peptides, including fusion proteins or peptides, encoded bynucleic acids as described herein.

In another aspect, the present invention provides host cells into whicha recombinant expression vector of the invention has been introduced. Ahost cell can be any prokaryotic cell (for example, E. coli) oreukaryotic cell (for example, yeast, insect, or mammalian cells (e.g.,CHO cells)). Vector DNA can be introduced into prokaryotic or eukaryoticcells via conventional transformation or transfection techniques. Forstable transfection of mammalian cells, it is known that, depending uponthe expression vector and transfection technique used, only a smallfraction of cells may integrate the foreign DNA into their genome. Inorder to identify and select these integrants, a gene that encodes aselectable marker (e.g., for resistance to antibiotics) is generallyintroduced into the host cells along with the gene of interest.Preferred selectable markers include those which confer resistance todrugs, such as G418, hygromycin and methotrexate. Cells stablytransfected with the introduced nucleic acid can be identified by drugselection (e.g., cells that have incorporated the selectable marker genewill survive, while the other cells die), among other methods.

Indications

TSLP is involved in promoting various inflammatory disorders, inparticular allergic inflammatory disorders. As used herein the term“allergic inflammation” refers to the manifestations of immunoglobulin E(IgE)-related immunological responses. (Manual of Allergy andImmunology, Chapter 2, Alvin M. Sanico, Bruce S. Bochner, and Sarbjit S.Saini, Adelman et al, ed., Lippincott, Williams, Wilkins, Philadelphia,Pa., (2002)). Allergic inflammation as used herein is generallycharacterized by the infiltration into the affected tissue of type 2helper T cells (T_(H)2 cells) (Kay, supra). Allergic inflammationincludes pulmonary inflammatory diseases such as allergicrhinosinusitis, asthma, allergic conjunctivitis, in addition toinflammatory skin conditions such as atopic dermatis (Manual of Allergyand Immunology, supra). As used herein the term “TSLP-related allergicinflammation” refers to allergic inflammation conditions in which TSLPis upregulated, or is demonstrated to be otherwise involved.

Allergic asthma is a chronic inflammatory disorder of the airwayscharacterized by airway eosinophilia, high levels of serum IgE and mastcell activation, which contribute to airway hyperresponsiveness,epithelial damage and mucus hypersecretion (Wills-Karp, M, Ann. Rev.Immunol. 17:255-281 (1999), Manual of Allergy and Immunology, supra).Studies have demonstrated that varying degrees of chronic inflammationare present in the airways of all asthmatics, even during symptom-freeperiods. In susceptible individuals, this inflammation causes recurrentepisodes of wheezing, breathlessness, chest tightness, and coughing.(Manual of Allergy and Immunology, supra).

Atopic dermatitis is a chronic pruritic inflammatory skin diseasecharacterized by skin lesions, featuring an elevated serum total IgE,eosinophilia, and increased release of histamine from basophils and mastcells. Persons suffering from atopic dermatitis exhibit exaggeratedT_(H)2 responses and initiation of atopic dermatitis lesions is thoughtto be mediated by means of early skin infiltration of T_(H)2 lymphocytesreleasing high levels of IL-4, IL-5 and IL-13 (Leung, J. Allergy ClinImmunol 105:860-76 (2000)). The relationship between TSLP and otherinflammatory cytokines is described in U.S. application Ser. No.11/205,904, publication 2006/0039910, which is herein incorporated byreference.

Human TSLP expression as detected by in situ hybridization was reportedto be increased in asthmatic airways correlating with disease severity(Ying et al., J. Immunology 174:8183-8190 (2005)). Analysis of TSLP mRNAlevels in asthmatic patient lung samples showed increased expression ofTSLP compared to controls. In addition, TSLP protein levels aredetectable in the concentrated bronchoalveolar lavage (BAL) fluid ofasthma patients, lung transplant patients, and cystic fibrosis patients.TSLP has recently been found to be released in response to microbes andtrauma as well as inflammation, and to activate mast cells (Allakhverdiet al., J. Exp. Med. 20492: 253-258 (2007).

Human TSLP protein was shown to correlate with disease in bronchialmucosa and BAL fluid of subjects with moderate/severe asthma and COPD.(Ying et al., J Immunol 181(4):2790-8 (2008).

Over-expression of TSLP in the lungs of transgenic mice leads toasthma-like airway inflammation (Zhou et al., Nat. Immunol 10:1047-1053(2005). In addition, it has been reported that TSLPR deficient micefailed to develop asthma in OVA-asthma models, demonstrating that TSLPis required for development of asthma in airway inflammation models(Zhou et al, supra, Carpino et al., Mol. Cell. Biol. 24:2584-2592(2004).

In addition to asthma, increased levels of TSLP protein and mRNA arefound in the lesional skin of atopic dermatitis (AD) patients and ininflamed tonsilar epithelial cells (Soumelis et al., Nature Immunol: 3(7): 673-680 (2002). Over-expression of TSLP in the skin of transgenicmice leads to an AD-like phenotype. (Yoo et al., J Exp Med 202:541-549(2005)).

Therefore, TSLP antagonists, specifically the TSLP antigen bindingproteins and antibodies of the instant application, are useful astherapeutic treatment for allergic inflammation, in particular, asthmaand atopic dermatitis.

In addition, TSLP antagonists, particularly the TSLP antigen bindingproteins and antibodies of the present disclosure are also useful fortreating fibrotic disorders. TSLP has been demonstrated to be involvedin promoting fibrotic disorders, as described in application Ser. No.11/344,379. TSLP has been found to induce fibroblast accumulation andcollagen deposition in animals. Injection of murine TSLP, for example,intradermally into mice resulted in fibrosis within the subcutis of themice, characterized by fibroblast proliferation and collagen deposition.Antagonizing TSLP activity would result in preventing or decreasingfibroblast proliferation and collagen deposition in a tissue.

As used herein the term “fibroproliferative disease” or “fibroticdisease or disorder” refers to conditions involving fibrosis in one ormore tissues. As used herein the term “fibrosis” refers to the formationof fibrous tissue as a reparative or reactive process, rather than as anormal constituent of an organ or tissue. Fibrosis is characterized byfibroblast accumulation and collagen deposition in excess of normaldeposition in any particular tissue. As used herein the term “fibrosis”is used synonymously with “fibroblast accumulation and collagendeposition”. Fibroblasts are connective tissue cells, which aredispersed in connective tissue throughout the body. Fibroblasts secretea nonrigid extracellular matrix containing type I and/or type IIIcollagen. In response to an injury to a tissue, nearby fibroblastsmigrate into the wound, proliferate, and produce large amounts ofcollagenous extracellular matrix. Collagen is a fibrous protein rich inglycine and proline that is a major component of the extracellularmatrix and connective tissue, cartilage, and bone. Collagen moleculesare triple-stranded helical structures called α-chains, which are woundaround each other in a ropelike helix. Collagen exists in several formsor types; of these, type I, the most common, is found in skin, tendon,and bone; and type III is found in skin, blood vessels, and internalorgans.

Fibrotic disorders include, but are not limited to, systemic and localscleroderma, keloids and hypertrophic scars, atherosclerosis,restenosis, pulmonary inflammation and fibrosis, idiopathic pulmonaryfibrosis, liver cirrhosis, fibrosis as a result of chronic hepatitis Bor C infection, kidney disease, heart disease resulting from scartissue, and eye diseases such as macular degeneration, and retinal andvitreal retinopathy. Additional fibrotic diseases include fibrosisresulting from chemotherapeutic drugs, radiation-induced fibrosis, andinjuries and burns.

Scleroderma is a fibrotic disorder characterized by a thickening andinduration of the skin caused by the overproduction of new collagen byfibroblasts in skin and other organs. Scleroderma may occur as a localor systemic disease. Systemic scleroderma may affect a number of organs.Systemic sclerosis is characterized by formation of hyalinized andthickened collagenous fibrous tissue, with thickening of the skin andadhesion to underlying tissues, especially of the hands and face. Thedisease may also be characterized by dysphagia due to loss ofperistalsis and submucosal fibrosis of the esophagus, dyspnea due topulmonary fibrosis, myocardial fibrosis, and renal vascular changes.(Stedman's Medical Dictionary, 26^(th) Edition, Williams & Wilkins,1995)). Pulmonary fibrosis affects 30 to 70% of scleroderma patients,often resulting in restrictive lung disease (Atamas et al. Cytokine andGrowth Factor Rev 14: 537-550 (2003)). Idiopathic pulmonary fibrosis isa chronic, progressive and usually lethal lung disorder, thought to be aconsequence of a chronic inflammatory process (Kelly et al., Curr PharmaDesign 9: 39-49 (2003)).

Therefore, TSLP antagonists, specifically the TSLP antigen bindingproteins and antibodies of the instant application, are useful astherapeutic treatment for fibrotic diseases, including but not limitedto scleroderma, interstitial lung disease, idiopathic pulmonaryfibrosis, fibrosis arising from chronic hepatitis B or C,radiation-induced fibrosis, and fibrosis arising from wound healing.

Although the above indications are preferred, other disease, disorder,or condition may be amenable to treatment with or may be prevented byadministration of an antigen binding to a subject. Such diseases,disorders, and conditions include, but are not limited to, inflammation,autoimmune disease, cartilage inflammation, fibrotic disease and/or bonedegradation, arthritis, rheumatoid arthritis, juvenile arthritis,juvenile rheumatoid arthritis, pauciarticular juvenile rheumatoidarthritis, polyarticular juvenile rheumatoid arthritis, systemic onsetjuvenile rheumatoid arthritis, juvenile ankylosing spondylitis, juvenileenteropathic arthritis, juvenile reactive arthritis, juvenile Reter'sSyndrome, SEA Syndrome (Seronegativity, Enthesopathy, ArthropathySyndrome), juvenile dermatomyositis, juvenile psoriatic arthritis,juvenile scleroderma, juvenile systemic lupus erythematosus, juvenilevasculitis, pauciarticular rheumatoid arthritis, polyarticularrheumatoid arthritis, systemic onset rheumatoid arthritis, ankylosingspondylitis, enteropathic arthritis, reactive arthritis, Reter'sSyndrome, SEA Syndrome (Seronegativity, Enthesopathy, ArthropathySyndrome), dermatomyositis, psoriatic arthritis, scleroderma, systemiclupus erythematosus, vasculitis, myolitis, polymyolitis,dermatomyolitis, osteoarthritis, polyarteritis nodossa, Wegener'sgranulomatosis, arteritis, ploymyalgia rheumatica, sarcoidosis,scleroderma, sclerosis, primary biliary sclerosis, sclerosingcholangitis, Sjogren's syndrome, psoriasis, plaque psoriasis, guttatepsoriasis, inverse psoriasis, pustular psoriasis, erythrodermicpsoriasis, dermatitis, atopic dermatitis, atherosclerosis, lupus,Still's disease, Systemic Lupus Erythematosus (SLE), myasthenia gravis,inflammatory bowel disease (IBD), Crohn's disease, ulcerative colitis,celiac disease, multiple schlerosis (MS), asthma, COPD, Guillain-Barredisease, Type I diabetes mellitus, Graves' disease, Addison's disease,Raynaud's phenomenon, autoimmune hepatitis, GVHD, and the like. Inspecific embodiments, pharmaceutical compositions comprising atherapeutically effective amount of TSLP antigen binding proteins areprovided.

The term “treatment” encompasses alleviation or prevention of at leastone symptom or other aspect of a disorder, or reduction of diseaseseverity, and the like. An antigen binding protein need not effect acomplete cure, or eradicate every symptom or manifestation of a disease,to constitute a viable therapeutic agent. As is recognized in thepertinent field, drugs employed as therapeutic agents may reduce theseverity of a given disease state, but need not abolish everymanifestation of the disease to be regarded as useful therapeuticagents. Similarly, a prophylactically administered treatment need not becompletely effective in preventing the onset of a condition in order toconstitute a viable prophylactic agent. Simply reducing the impact of adisease (for example, by reducing the number or severity of itssymptoms, or by increasing the effectiveness of another treatment, or byproducing another beneficial effect), or reducing the likelihood thatthe disease will occur or worsen in a subject, is sufficient. Oneembodiment of the invention is directed to a method comprisingadministering to a patient an antigen binding protein in an amount andfor a time sufficient to induce a sustained improvement over baseline ofan indicator that reflects the severity of the particular disorder.

Pharmaceutical Compositions

In some embodiments, the invention provides pharmaceutical compositionscomprising a therapeutically effective amount of one or a plurality ofthe antigen binding proteins of the invention together with apharmaceutically acceptable diluent, carrier, solubilizer, emulsifier,preservative, and/or adjuvant. In addition, the invention providesmethods of treating a patient by administering such pharmaceuticalcomposition. The term “patient” includes human and animal subjects.

Pharmaceutical compositions comprising one or more antigen bindingproteins may be used to reduce TSLP activity. Pharmaceuticalcompositions comprising one or more antigen binding proteins may be usedin treating the consequences, symptoms, and/or the pathology associatedwith TSLP activity. Pharmaceutical compositions comprising one or moreantigen binding proteins may be used in methods of inhibiting bindingand/or signaling of TSLP to TSLPR comprising providing the antigenbinding protein of the invention to TSLP.

In certain embodiments, acceptable formulation materials preferably arenontoxic to recipients at the dosages and concentrations employed. Incertain embodiments, the pharmaceutical composition may containformulation materials for modifying, maintaining or preserving, forexample, the pH, osmolality, viscosity, clarity, color, isotonicity,odor, sterility, stability, rate of dissolution or release, adsorptionor penetration of the composition. In such embodiments, suitableformulation materials include, but are not limited to, amino acids (suchas glycine, glutamine, asparagine, arginine or lysine); antimicrobials;antioxidants (such as ascorbic acid, sodium sulfite or sodiumhydrogen-sulfite); buffers (such as borate, bicarbonate, Tris-HCl,citrates, phosphates or other organic acids); bulking agents (such asmannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine,polyvinylpyrrolidone, beta-cyclodextrin orhydroxypropyl-beta-cyclodextrin); fillers; monosaccharides;disaccharides; and other carbohydrates (such as glucose, sucrose,mannose or dextrins); proteins (such as serum albumin, gelatin orimmunoglobulins); coloring, flavoring and diluting agents; emulsifyingagents; hydrophilic polymers (such as polyvinylpyrrolidone); lowmolecular weight polypeptides; salt-forming counterions (such assodium); preservatives (such as benzalkonium chloride, benzoic acid,salicylic acid, thimerosal, phenethyl alcohol, methylparaben,propylparaben, chlorhexidine, sorbic acid or hydrogen peroxide);solvents (such as glycerin, propylene glycol or polyethylene glycol);sugar alcohols (such as mannitol or sorbitol); suspending agents;surfactants or wetting agents (such as pluronics, PEG, sorbitan esters,polysorbates such as polysorbate 20, polysorbate, triton, tromethamine,lecithin, cholesterol, tyloxapal); stability enhancing agents (such assucrose or sorbitol); tonicity enhancing agents (such as alkali metalhalides, preferably sodium or potassium chloride, mannitol sorbitol);delivery vehicles; diluents; excipients and/or pharmaceutical adjuvants.See, REMINGTON'S PHARMACEUTICAL SCIENCES, 18″ Edition, (A. R. Genrmo,ed.), 1990, Mack Publishing Company.

In certain embodiments, the optimal pharmaceutical composition will bedetermined by one skilled in the art depending upon, for example, theintended route of administration, delivery format and desired dosage.See, for example, REMINGTON'S PHARMACEUTICAL SCIENCES, supra. In certainembodiments, such compositions may influence the physical state,stability, rate of in vivo release and rate of in vivo clearance of theantigen binding proteins of the invention. In certain embodiments, theprimary vehicle or carrier in a pharmaceutical composition may be eitheraqueous or non-aqueous in nature. For example, a suitable vehicle orcarrier may be water for injection, physiological saline solution orartificial cerebrospinal fluid, possibly supplemented with othermaterials common in compositions for parenteral administration. Neutralbuffered saline or saline mixed with serum albumin are further exemplaryvehicles. In specific embodiments, pharmaceutical compositions compriseTris buffer of about pH 7.0-8.5, or acetate buffer of about pH 4.0-5.5,and may further include sorbitol or a suitable substitute therefor. Incertain embodiments of the invention, TSLP antigen binding proteincompositions may be prepared for storage by mixing the selectedcomposition having the desired degree of purity with optionalformulation agents (REMINGTON'S PHARMACEUTICAL SCIENCES, supra) in theform of a lyophilized cake or an aqueous solution. Further, in certainembodiments, the TSLP antigen binding protein product may be formulatedas a lyophilizate using appropriate excipients such as sucrose.

The pharmaceutical compositions of the invention can be selected forparenteral delivery. Alternatively, the compositions may be selected forinhalation or for delivery through the digestive tract, such as orally.The formulation components are present preferably in concentrations thatare acceptable to the site of administration. In certain embodiments,buffers are used to maintain the composition at physiological pH or at aslightly lower pH, typically within a pH range of from about 5 to about8. Including about 5.1, about 5.2, about 5.3, about 5.4, about 5.5,about 5.6, about 5.7, about 5.8, about 5.9, about 6.0, about 6.1, about6.2, about 6.3, about 6.4, about 6.5, about 6.6, about 6.7, about 6.8,about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about7.5, about 7.6, about 7.7, about 7.8, about 7.9, and about 8.0.

When parenteral administration is contemplated, the therapeuticcompositions for use in this invention may be provided in the form of apyrogen-free, parenterally acceptable aqueous solution comprising thedesired TSLP antigen binding protein in a pharmaceutically acceptablevehicle. A particularly suitable vehicle for parenteral injection issterile distilled water in which the TSLP antigen binding protein isformulated as a sterile, isotonic solution, properly preserved. Incertain embodiments, the preparation can involve the formulation of thedesired molecule with an agent, such as injectable microspheres,bio-erodible particles, polymeric compounds (such as polylactic acid orpolyglycolic acid), beads or liposomes, that may provide controlled orsustained release of the product which can be delivered via depotinjection. In certain embodiments, hyaluronic acid may also be used,having the effect of promoting sustained duration in the circulation. Incertain embodiments, implantable drug delivery devices may be used tointroduce the desired antigen binding protein.

Pharmaceutical compositions of the invention can be formulated forinhalation. In these embodiments, TSLP antigen binding proteins areadvantageously formulated as a dry, inhalable powder. In specificembodiments, TSLP antigen binding protein inhalation solutions may alsobe formulated with a propellant for aerosol delivery. In certainembodiments, solutions may be nebulized. Pulmonary administration andformulation methods therefore are further described in InternationalPatent Application No. PCTUS94/001875, which is incorporated byreference and describes pulmonary delivery of chemically modifiedproteins.

It is also contemplated that formulations can be administered orally.TSLP antigen binding proteins that are administered in this fashion canbe formulated with or without carriers customarily used in thecompounding of solid dosage forms such as tablets and capsules. Incertain embodiments, a capsule maybe designed to release the activeportion of the formulation at the point in the gastrointestinal tractwhen bioavailability is maximized and pre-systemic degradation isminimized. Additional agents can be included to facilitate absorption ofthe TSLP antigen binding protein. Diluents, flavorings, low meltingpoint waxes, vegetable oils, lubricants, suspending agents, tabletdisintegrating agents, and binders may also be employed.

A pharmaceutical composition of the invention is preferably provided tocomprise an effective quantity of one or a plurality of TSLP antigenbinding proteins in a mixture with non-toxic excipients that aresuitable for the manufacture of tablets. By dissolving the tablets insterile water, or another appropriate vehicle, solutions may be preparedin unit-dose form.

Suitable excipients include, but are not limited to, inert diluents,such as calcium carbonate, sodium carbonate or bicarbonate, lactose, orcalcium phosphate; or binding agents, such as starch, gelatin, oracacia; or lubricating agents such as magnesium stearate, stearic acid,or talc.

Additional pharmaceutical compositions will be evident to those skilledin the art, including formulations involving TSLP antigen bindingproteins in sustained- or controlled-delivery formulations. Techniquesfor formulating a variety of other sustained- or controlled-deliverymeans, such as liposome carriers, bio-erodible microparticles or porousbeads and depot injections, are also known to those skilled in the art.See, for example, International Patent Application No. PCT/US93/00829,which is incorporated by reference and describes controlled release ofporous polymeric microparticles for delivery of pharmaceuticalcompositions.

Sustained-release preparations may include semipermeable polymermatrices in the form of shaped articles, e.g., films, or microcapsules.Sustained release matrices may include polyesters, hydrogels,polylactides (as disclosed in U.S. Pat. No. 3,773,919 and EuropeanPatent Application Publication No. EP 058481, each of which isincorporated by reference), copolymers of L-glutamic acid and gammaethyl-L-glutamate (Sidman et al, 1983, Biopolymers 2:547-556), poly(2-hydroxyethyl-inethacrylate) (Langer et al, 1981, J. Biomed. Mater.Res. 15:167-277 and Langer, 1982, Chem. Tech. 12:98-105), ethylene vinylacetate (Langer et al, 1981, supra) or poly-D(−)-3-hydroxybutyric acid(European Patent Application Publication No. EP 133,988).

Sustained release compositions may also include liposomes that can beprepared by any of several methods known in the art. See, e.g., Eppsteinet al, 1985, Proc. Natl. Acad. ScL U.S.A. 82.3688-3692; European PatentApplication Publication Nos. EP 036,676; EP 088,046 and EP 143,949,incorporated by reference.

Pharmaceutical compositions used for in vivo administration aretypically provided as sterile preparations. Sterilization can beaccomplished by filtration through sterile filtration membranes. Whenthe composition is lyophilized, sterilization using this method may beconducted either prior to or following lyophilization andreconstitution. Compositions for parenteral administration can be storedin lyophilized form or in a solution. Parenteral compositions generallyare placed into a container having a sterile access port, for example,an intravenous solution bag or vial having a stopper pierceable by ahypodermic injection needle.

Aspects of the invention includes self-buffering TSLP antigen bindingprotein formulations, which can be used as pharmaceutical compositions,as described in international patent application WO 0613818 1A2(PCT/US2006/022599), which is incorporated by reference in its entiretyherein. One embodiment provides self-buffering TSLP antigen bindingprotein formulations comprising an TSLP antigen binding protein in whichthe total salt concentration is less than 150 mM.

The therapeutically effective amount of TSLP antigen bindingprotein-containing pharmaceutical composition to be employed willdepend, for example, upon the therapeutic context and objectives. Oneskilled in the art will appreciate that the appropriate dosage levelsfor treatment will vary depending, in part, upon the molecule delivered,the indication for which the TSLP antigen binding protein is being used,the route of administration, and the size (body weight, body surface ororgan size) and/or condition (the age and general health) of thepatient.

In certain embodiments, the clinician may titer the dosage and modifythe route of administration to obtain the optimal therapeutic effect. Atypical dosage may range from about 0.1 μg/kg to up to about 30 mg/kg ormore, depending on the factors mentioned above. In specific embodiments,the dosage may range from 0.1 μg/kg up to about 30 mg/kg, optionallyfrom 1 μg/kg up to about 30 mg/kg or from 10 μg/kg up to about 5 mg/kg.

Dosing frequency will depend upon the pharmacokinetic parameters of theparticular TSLP antigen binding protein in the formulation used.Typically, a clinician administers the composition until a dosage isreached that achieves the desired effect. The composition may thereforebe administered as a single dose, or as two or more doses (which may ormay not contain the same amount of the desired molecule) over time, oras a continuous infusion via an implantation device or catheter. Furtherrefinement of the appropriate dosage is routinely made by those ofordinary skill in the art and is within the ambit of tasks routinelyperformed by them.

Appropriate dosages may be ascertained through use of appropriatedose-response data. In certain embodiments, the antigen binding proteinsof the invention can be administered to patients throughout an extendedtime period. Chronic administration of an antigen binding protein of theinvention minimizes the adverse immune or allergic response commonlyassociated with antigen binding proteins that are not fully human, forexample an antibody raised against a human antigen in a non-humananimal, for example, a non-fully human antibody or non-human antibodyproduced in a non-human species.

The route of administration of the pharmaceutical composition is inaccord with known methods, e.g., orally, through injection byintravenous, intraperitoneal, intracerebral (intra-parenchymal),intracerebroventricular, intramuscular, intra-ocular, intraarterial,intraportal, or intralesional routes; by sustained release systems or byimplantation devices. In certain embodiments, the compositions may beadministered by bolus injection or continuously by infusion, or byimplantation device.

The composition also may be administered locally via implantation of amembrane, sponge or another appropriate material onto which the desiredmolecule has been absorbed or encapsulated. In certain embodiments,where an implantation device is used, the device may be implanted intoany suitable tissue or organ, and delivery of the desired molecule maybe via diffusion, timed-release bolus, or continuous administration.

Combination Therapies

In further embodiments, antigen binding protein are administered incombination with other agents useful for treating the condition withwhich the patient is afflicted. Examples of such agents include bothproteinaceous and non-proteinaceous drugs. When multiple therapeuticsare co-administered, dosages may be adjusted accordingly, as isrecognized in the pertinent art. “Co-administration” and combinationtherapy are not limited to simultaneous administration, but also includetreatment regimens in which an antigen binding protein is administeredat least once during a course of treatment that involves administeringat least one other therapeutic agent to the patient.

The invention having been described, the following examples are offeredby way of illustration, and not limitation.

EXAMPLE 1 Preparation of Antigen

Several forms of recombinant TSLP were used as immunogens. Human TSLPwas expressed both in E. coli and in mammalian cells. The E. coliproduced human TSLP was an untagged full-length protein. TSLP proteinwas produced in COS PKB cells having a deleted furin cleavage siteproduced by deleting nucleotides 382-396 (AGAAAAAGGAAAGTC, SEQ ID NO:370) corresponding to amino acids 128-132 (RKRKV, SEQ ID NO: 371). Thisprotein contained a C terminal polyHIS-Flag tag (Nucleotidesequence=ATGTTCCCTTTTGCCTTACTATATGTTCTGTCAGTTTCTTTCAGGAAAATCTTCATCTTACAACTTGTAGGGCTGGTGTTAACTTACGACTTCACTAACTGTGACTTTGAGAAGATTAAAGCAGCCTATCTCAGTACTATTTCTAAAGACCTGATTACATATATGAGTGGGACCAAAAGTACCGAGTTCAACAACACCGTCTCTTGTAGCAATCGGCCACATTGCCTTACTGAAATCCAGAGCCTAACCTTCAATCCCACCGCCGGCTGCGCGTCGCTCGCCAAAGAAATGTTCGCCATGAAAACTAAGGCTGCCTTAGCTATCTGGTGCCCAGGCTATTCGGAAACTCAGATAAATGCTACTCAGGCAATGAAGAAGAGGACAACCAATAAATGTCTGGAACAAGTGTCACAATTACAAGGATTGTGGCGTCGCTTCAATCGACCTTTACTGAAACAACAGCATCACCATCACCATCACGACTACAAAGACGATGACGACAAA (SEQ ID NO: 372);

Proteinsequence=MFPFALLYVLSVSFRKIFILQLVGLVLTYDFTNCDFEKIKAAYLSTISKDLITYMSGTKSTEFNNTVSCSNRPHCLTEIQSLTFNPTAGCASLAKEMFAMKTKAALAIWCPGYSETQINATQAMKKRTTNKCLEQVSQLQGLWRRFNRPLLKQQHHHHHHDYKDDDDK (SEQ ID NO: 373).

In another campaign, a full length TSLP C terminal polyHIS-Flag taggedprotein was produced in COS PKB cells (Nucleotidesequence=ATGTTCCCTTTTGCCTTACTATATGTTCTGTCAGTTTCTTTCAGGAAAATCTTCATCTTACAACTTGTAGGGCTGGTGTTAACTTACGACTTCACTAACTGTGACTTTGAGAAGATTAAAGCAGCCTATCTCAGTACTATTTCTAAAGACCTGATTACATATATGAGTGGGACCAAAAGTACCGAGTTCAACAACACCGTCTCTTGTAGCAATCGGCCACATTGCCTTACTGAAATCCAGAGCCTAACCTTCAATCCCACCGCCGGCTGCGCGTCGCTCGCCAAAGAAATGTTCGCCATGAAAACTAAGGCTGCCTTAGCTATCTGGTGCCCAGGCTATTCGGAAACTCAGATAAATGCTACTCAGGCAATGAAGAAGAGGAGAAAAAGGAAAGTCACAACCAATAAATGTCTGGAACAAGTGTCACAATTACAAGGATTGTGGCGTCGCTTCAATCGACCTTTACTGAAACAACAGCATCACCATCACCATCACGACTACAAAGACGATGACGACAAA (SEQ ID NO: 374); Proteinsequence=MFPFALLYVLSVSFRKIFILQLVGLVLTYDFTNCDFEKIKAAYLSTISKDLITYMSGTKSTEFNNTVSCSNRPHCLTEIQSLTFNPTAGCASLAKEMFAMKTKAALAIWCPGYSETQINATQAMKKRRKRKVTTNKCLEQVSQLQGLWRRFNRPLLKQQHHHHHHDYKDDDDK (SEQ ID NO: 375). Notethat the amino acid sequence 1-28 (MFPFALLYVLSVSFRKIFLQLVGLVLT, SEQ IDNO: 376) is a signal peptide cleaved from the mature product of boththese proteins.

In addition, cynomolgus TSLP was cloned and subcloned/expressedsimilarly with either the furin cleavage site (nucleotide 358-372(AGAAAAAGGAAAGTC, SEQ ID NO: 370) corresponding to amino acids 120-124(RKRKV, SEQ ID NO: 371)) deleted(DNA=ATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACCGGTTACGACTTCACTAACTGTGACTTTCAGAAGATTGAAGCAGACTATCTCCGTACTATTTCTAAAGACCTGATTACATATATGAGTGGGACTAAAAGTACCGACTTCAACAACACCGTCTCCTGTAGCAATCGGCCACACTGCCTTACTGAAATCCAGAGCCTAACCTTCAATCCCACCCCCCGCTGCGCGTCGCTCGCCAAGGAAATGTTCGCCAGGAAAACTAAGGCTACCCTCGCTCTCTGGTGCCCAGGCTATTCGGAAACTCAGATAAATGCTACTCAGGCAATGAAGAAGAGGACAACCAATAAATGTCTGGAACAAGTGTCACAATTACTAGGATTGTGGCGTCGCTTCATTCGAACTTTACTGAAACAACAGCACCACCACCACCACCATGACTATAAAGACGATGACGAC AAAT (SEQ IDNO: 377);Protein=METDTLLLWVLLLWVPGSTGYDFTNCDFQKIEADYLRTISKDLITYMSGTKSTDFNNTVSCSNRPHCLTEIQSLTFNPTPRCASLAKEMFARKTKATLALWCPGYSETQINATQAMKKRTTNKCLEQVSQLLGLWRRFIRTLLKQQHHHHHHDYKDDDDK (SEQ ID NO: 378) or as afull-length/native product (nucleotidesequence=ATGGAGACAGACACACTCCTGCTATGGGTACTGCTGCTCTGGGTTCCAGGTTCCACCGGTTACGACTTCACTAACTGTGACTTTCAGAAGATTGAAGCAGACTATCTCCGTACTATTTCTAAAGACCTGATTACATATATGAGTGGGACTAAAAGTACCGACTTCAACAACACCGTCTCCTGTAGCAATCGGCCACACTGCCTTACTGAAATCCAGAGCCTAACCTTCAATCCCACCCCCCGCTGCGCGTCGCTCGCCAAGGAAATGTTCGCCAGGAAAACTAAGGCTACCCTCGCTCTCTGGTGCCCAGGCTATTCGGAAACTCAGATAAATGCTACTCAGGCAATGAAGAAGAGGAGAAAAAGGAAAGTCACAACCAATAAATGTCTGGAACAAGTGTCACAATTACTAGGATTGTGGCGTCGCTTCATTCGAACTTTACTGAAACAACAGCACCACCACCACCACCATGACTATAAAGACGATGACGACAAA (SEQ ID NO: 379);Protein=METDTLLLWVLLLWVPGSTGYDFTNCDFQKIEADYLRTISKDLITYMSGTKSTDFNNTVSCSNRPHCLTEIQSLTFNPTPRCASLAKEMFARKTKATLALWCPGYSETQINATQAMKKRRKRKVTTNKCLEQVSQLLGLWRRFIRTLLKQQHHHHHHDYKDDDDK (SEQ ID NO: 380) fused to thesame C terminal polyHIS-Flag in COS PKB cells. Note that the amino acidsequence 1-20 (METDTLLLWVLLLWVPGSTG, SEQ ID NO: 381) is a signal peptidecleaved from the mature product of both these cynomolgus proteins.

EXAMPLE 2 Mouse Anti-Human TSLP Antibodies

hTSLP-Fc was used for immunization of Balb/c mice (Jackson Laboratories,Bar Harbor, Me.). After several rounds of immunization, lymphocytes werereleased from the spleen and were fused with mouse myeloma cells, NS1(ATCC) by chemical fusion with 50% PEG/DMSO (Sigma). The fused cellswere seeded in 96-well plates at the density of 2×10⁴ cells/well in 200ul of DMEM HAT (0.1 mM hypoxanthine, 0.16 mM thymidine, 4 mMaminopterin, Sigma) media supplemented with 10% FBS, 5% Origen CloningFactor (BioVeris™), 1× Penicillin-Streptomycin-Glutamine, SodiumPyruvate (Invitrogen). Medium was replaced 7 days post-fusion with DMEMHT (0.1mM hypoxanthine, 0.16 mM thymidine) media supplemented with 10%FBS, 5% Origen Cloning Factor (BioVeris™), 1×Penicillin-Streptomycin-Glutamine, Sodium Pyruvate (Invitrogen).Conditioned media was collected two days after medium change andpreceded for primary screening.

EXAMPLE 3 Fully Human Antibody Generation

Fully human monoclonal antibodies specific for TSLP were generated usingthe XenoMouse® technology according to protocols described, for example,in U.S. 2005/0118643, U.S. Pat. Nos. 6,114,598, 6,162,963, 7,049,426,7,064,244, Green et al., Nature Genetics 7:13-21 (1994), Medez et al.Nature Genetics 15:146-156 (1997), Green and Jakobovitis J. Ex. Med.188:483-495 (1998) (all of which are incorporated by reference herein),and as described below.

Two campaigns were conducted. In campaign 1, IgG2 and IgG4 cohorts ofXenoMouse® were utilized. 50% of the mice received E. coli producedhuman TSLP and 50% received mammalian produced human TSLP (describedabove). Serum titers were monitored by ELISA (described below) and micewith the best titers were fused to generate hybridomas using thefollowing protocols.

Selected mice were sacrificed and the draining lymph nodes harvested andpooled from each cohort. The lymphoid cells were enriched for B cellsand the B cells fused with myeloma cells to create hybridomas. The fusedhybridoma lines were then plated in hybridoma media and cultured for10-14 days at 37° C. The hybridoma supernatants were screened for IgGantibodies binding to TSLP by ELISA as described below.

A second campaign was initiated in which two cohorts of IgG2 XenoMouse®were immunized with mammalian produced human TSLP, and one cohort wasboosted with cynomolgus TSLP. After several rounds of immunization,lymphocytes from lymph nodes were fused and cultured as described above.After culturing, hybridoma supernatants were screened for binding toTSLP by ELISA, as described below.

The polyclonal supernatants from both campaigns were selected forfurther subcloning on the basis of the assays set out below. Thehybridomas containing antibodies that are potent inhibitors of TSLPactivity were identified, and cross-reactivity with cyno TSLP wasfurther determined. The results are shown in Example 5 below. Promisinghybridoma supernatants were selected on the basis of their performancein the primary DC assay described below. Those hybridomas were singlecell cloned and expanded for further testing. The antibodies were thenpurified as described below.

Antibodies were purified from conditioned media of the hybridomas usingMab Select (GE Healthcare) resin. 100 ul of a 1:2 slurry of Mab Selectresin equilibrated in PBS was added to between 7 and 10 ml ofconditioned media (CM). The tubes were placed on rotators at 4-8° C.overnight. The tubes were centrifuged at 1,000×g for 5 minutes and thenon-bound fraction was decanted. The resin was washed with 5 ml of PBS,and centrifuged and decanted as above. The resin was then transferred toa SPIN-X, 0.45 um, 2 ml tube. The resin was washed an additional twotimes with 0.5 ml of PBS and centrifuged. The Mabs were eluted with 0.2ml of 0.1M acetic acid by incubating at room temperature with occasionalmixing for 10 minutes. The tubes were centrifuged, and 30 ul of 1M Trisbuffer Ph 8.0 is added to the eluate. Purified Mab's were stored 4-8° C.

EXAMPLE 4 Antibody Assays

A. ELISA to Detect Presence of Anti-TSLP Antibody

ELISAs were performed by coating Costar 3368 medium binding 96 wellplates with recombinantly produced wtHuTSLP or pHisFlag at 2 ug/ml 50ul/well in 1×PBS/0.05% azide, and incubated overnight at 4° C. Theplates were washed and blocked with 250 ul of 1×PBS/1% milk (the assaydiluent), and incubated at least 30 minutes at room temperature.

Approximately 50 ul/well hybridoma supernatants, positive control mouseantibody M385, or negative control were added, and incubated at roomtemperature for 2 hours. The plates were washed, and a secondaryantibody, goat anti-human IgG Fc HPR (Pierce), or alternatively a goatanti-mouse IgG HPR (Jackson Labs), was applied at 400 ng/ml in assaydiluent. The plates were incubated 1 hr at RT, washed, and the OD at 450nm read.

B. Screening of Anti-TSLP Hybridoma Supernatants was Performed Using Oneof the Following Functional Assays

1. 96 well plates were coated with soluble huIL-7Ra-huTSLPR-Fc protein,with an 8 aa acid linker (SGGAPMLS, SEQ ID NO: 382) between the receptorand a human Fc, and incubated overnight at 4° C.

2. The plates were washed and blocked for 1 hour at RT with PBS+1%BSA+5% sucrose.

3. The plates were incubated with biotinylated huTSLPHFdel (HF standsfor polyHis Flag, where the TSLP has the furin cleavage site deleted)(del). The plates were then incubated (+/−) hybridoma supernatants ormouse anti-human TSLP (M385) as a positive control for 2 h at RT.

4. SA-HRP detection (streptavidin-horseradish peroxidase). SA bindsstrongly to the biotin portion of biotinylated huTSLPHFdel and HRPcatalyzes the oxidation of the chromogen, TMB (which turns blue), byhydrogen peroxide.

B. Cell Based Assays

1) The inhibition of TSLP-induced proliferation of stable BAF cell lineexpressing the human TSLPR-IL7R complex by hybridoma supernatants orpurified antibodies was determined according to the following protocol.

1. BAF: Hu TSLPR stable cell lines in growth media, RPMI 1640+10% FBS+1%L-Glutamine+0.1% Pen/Strep +0.1% 2-ME were washed to remove TSLP used inmaintenance media, that is the same as the growth media but with theaddition of 10 ng/mL of huTSLPHFwt.

2. HuTSLPwtpHF (+/−) or cynomolgus TSLPwtpHF (+/−) were incubated withhybridoma supernatants/purified antibody/or mouse anti-human TSLP (M385)for 30 minutes at room temperature in wells.

3. 5×10⁴ BAF cells/well were added and incubated for 3 days.

4. The cells were pulsed with tritiated thymidine (1 uCi/well)overnight. Cell proliferation of the BAF cells, or the inhibitionthereof, was assessed by the amount of tritiated thymidine incorporation(CPM) by the cells.

2) Primary cell assay. Inhibition of TSLP induced osteoprotegerin (OPG)(described in U.S. Pat. No. 6,284,728) production from primary humandendritic cells (DC) by hybridomas or purified antibodies was determinedaccording to the following protocol.

1. Peripheral blood CD11c+ myeloid DCs were enriched from normal inhousedonor leukapheresis packs using CD1c(BDCA-1) DC isolation kit (MiltenyiBiotec).

2. huTSLPwtpHF (+/−) or cynomolgus TSLPwtpHF were incubated withsupernatants or purified antibody or mouse anti-human TSLP for 30minutes at room temperature.

3. 1×105 cells/well were added and incubated for 48 hours. Supernatantswere harvested and assayed for human OPG production by ELISA, and theinhibition of OPG production by the hybridoma supernatants or purifiedantibodies was determined. The OPG ELISA was performed using an R&Dsystems DuoSet® development kit. Anti-TSLP antibodies inhibited OPGproduction from cells in a dose-dependent manner.

3) Cynomolgus Peripheral Blood Mononuclear Cell Assay. Inhibition ofCynoTSLP induced CCL22/MDC production by hybridoma supernatants orpurified antibodies was determined according to the following protocol.

1. Peripheral blood mononuclear cells (PBMC) from peripheral bloodobtained from cynomolgus monkeys (SNBL) were obtained by overlaying 1:1blood:PBS mixture over isolymph.

2. Cynomolgus TSLPwtpHF(+/−) supernatants/purified antibody or solublehuIL-7Ra-huTSLPR-Fc were incubated for 30 minutes at room temperature.

3. 4×10⁵ cells/well were added and incubated for 5 days. Thesupernatants were harvested and assayed for cynomolgus CCL22/MDCproduction by ELISA.

EXAMPLE 5 K_(D) Determinations

The surface plasmon resonance experiments described in this patentapplication were conducted at 25° C. using a Biacore 3000 instrument(Biacore International AB, Uppsala, Sweden) equipped with a CM4 sensorchip. Anti-Fcγ specific capture antibodies were covalently immobilizedto two flow cells on the CM4 chip using standard amine-couplingchemistry with HBS-EP as the running buffer. Briefly, each flow cell wasactivated with a 1:1 (v/v) mixture of 0.1 M NHS and 0.4 M EDC.AffiniPure Goat Anti-Human IgG, Fcγ Fragment Specific antibody (JacksonImmunoResearch Inc. West Grove, Pa.) at 30 ug/ml in 10 mM sodiumacetate, pH 5.0 was immobilized with a target level of 3,000 RUs on twoflow cells. Residual reactive surfaces were deactivated with aninjection of 1 M ethanolamine. The running buffer was then switched toHBS-EP+0.1 mg/ml BSA for all remaining steps.

The following antibodies were tested. A5 IgG2 was a purified clonalantibody, A2 IgG1 and IgG2 were recombinant purified antibodies, and A3IgG4 and A4 IgG4 were clonal supernatants. The antibodies were dilutedappropriately in running buffer so that a 2 minute injection at 10μl/min over the test flow cell resulted in approximately 110-175response units of antibody captured on the test flow cell surface. Noantibody was captured on the control flow cell surface. Human, cyno, ormurine TSLP at various concentrations, along with buffer blanks werethen flown over the two flow cells. The concentration ranges for humanand cyno TSLP were from 0.44-100 nM while the concentration range formurine TSLP was from 8.2-6000 nM. A flow rate of 50 ul/min was used anda 2 minute association phase followed by a 10-30 minute dissociationphase. After each cycle the surfaces were regenerated with a 30 secondinjection of 10 mM glycine pH 1.5. Fresh antibody was then captured onthe test flow cell to prepare for the next cycle.

Data was double referenced by subtracting the control surface responsesto remove bulk refractive index changes, and then subtracting theaveraged buffer blank response to remove systematic artifacts from theexperimental flow cells. The TSLP data were processed and globally fitto a 1:1 interaction model with a local Rmax in BIA evaluation Softwarev 4.1. (Biacore International AB, Uppsala, Sweden). Association (k_(a))and dissociation (k_(d)) rate constants were determined and used tocalculate the dissociation equilibrium constant (K_(D)). Thedissociation rate constants and dissociation equilibrium constants aresummarized in the table found in Example 6.

EXAMPLE 6 In Vitro Activity of Antibodies

The following antibodies were characterized using the Biacore assaydescribed above for kd and KD. The primary dendritic cell assay was usedfor determining IC₅₀ (pM). The data for A5 was generated with purifiedclonal antibody, for A2 was generated with recombinant purifiedantibody, and data for A3 and A4 was generated using clonal supernatant.All versions of TSLP were generated from mammalian cells.

Antibody TSLP kd (1/x) off-rate KD (pM) IC50 (pM) A5 IgG2 Hu TSLP 7.36 ×10⁻⁵ 29.2 100-220 Cyno TSLP 8.64 × 10⁻⁵ 51.2 680-970 Mu TSLP 8.81 × 10⁻⁴377,000 Nd A2 IgG1 Hu TSLP 3.49 × 10⁻⁴ 203 600-1700 Cyno TSLP 1.04 ×10⁻⁴ 46.8 250-860 Mu TSLP — — — A2 IgG2 Hu TSLP 2.85 × 10⁻⁴ 157  6-24Cyno TSLP 9.42 × 10⁻⁵ 37.6 Nd Mu TSLP no binding no binding n/a A3 IgG4Hu TSLP 2.7 × 10⁻⁴ 170  6-24 Cyno TSLP Nd nd Nd Mu TSLP Nd nd Nd A4 IgG4Hu TSLP 3.30 × 10⁻⁴ 340 30-59 Cyno TSLP Nd nd Nd Mu TSLP Nd nd Nd

EXAMPLE 7 Recombinant Expression and Purification of AntibodiesDevelopment of Stable Cell Line Expressing Antibodies

Overlapping oligonucleotides were synthesized corresponding to theprimary sequence of the light chain or heavy chain variable domain forboth the sense and anti-sense strand. This oligonucleotide pool wasemployed in a standard PCR. Product from this first reaction was used astemplate in a second PCR amplification. Amplified variable heavy chainand variable light chain fragments were sub-cloned into an intermediatevector and sequenced to identify error-free products. The variable heavychain fragment was cloned into a transient expression vector containinga signal peptide and human IgG2 constant region. The variable lightchain fragment was cloned into a transient expression vector containinga signal peptide and human lambda constant region. The complete heavychain gene was transferred into the vector pDC324. The complete lightchain gene was transferred into the expression vector, pDC323.

The CS-9 host cells used for transfection of the anti-TSLP expressionplasmids are a CHO cell line derived from DXB-11 cells throughadaptation to serum-free media (Rasmussen et al, Cytotechnology28:31-42, 1998). The anti-TSLP cell lines were created by transfectingCS-9 host cells with the expression plasmids pDC323-anti-TSLP-lambda andpDC324-anti-TSLP-IgG2 using a standard electroporation or lipofectionprocedure. After transfection of the host cell line with the expressionplasmids, the cells were grown in selection medium for 2-3 weeks toallow for selection of the plasmids and recovery of the cells. In somecases, the medium was supplemented with 3% dialyzed fetal bovine serum(ds or dFBS). If serum was used, it was removed from the medium afterthe selection period. The cells were grown in selective medium untilthey achieved >85% viability. This pool of transfected cells was thencultured in culture medium.

Cell Line Cloning

A cell bank was made of selected clones according to the followingprocedure. The cloning step ensures that clonal populations and cellbanks were generated enabling a reproducible performance in commercialmanufacturing. An amplified pool of antibody-expressing cells was seededunder limiting dilution in 96-well plates, and candidate clones wereevaluated for growth and productivity performance in small-scale studies

EXAMPLE 8 Antibody Cross-Competition

A common way to define epitopes is through competition experiments.Antibodies that compete with each other can be thought of as binding thesame site on the target. This example describes a method of determiningcompetition for binding to TSLP and the results of the method whenapplied to a number of antibodies described herein.

Binning experiments can be conducted in a number of ways, and the methodemployed may have an effect on the assay results. Common to thesemethods is that TSLP is typically bound by one reference antibody andprobed by another. If the reference antibody prevents the binding of theprobe antibody then the antibodies are said to be in the same bin. Theorder in which the antibodies are employed is important. If antibody Ais employed as the reference antibody and blocks the binding of antibodyB the converse is not always true: antibody B used as the referenceantibody will not necessarily block antibody A. There are a number offactors in play here: the binding of an antibody can causeconformational changes in the target which prevent the binding of thesecond antibody, or epitopes which overlap but do not completely occludeeach other may allow for the second antibody to still have enoughhigh-affinity interactions with the target to allow binding. Antibodieswith a much higher affinity may have a greater ability to bump ablocking antibody out of the way. In general, if competition is observedin either order the antibodies are said to bin together, and if bothantibodies can block each other then it is likely that the epitopesoverlap more completely.

For this Example, a modification of the Multiplexed Binning methoddescribed by Jia, et al (J. Immunological Methods, 288 (2004) 91-98) wasused. Because the presence of a furin cleavage site within TSLP can leadto heterogeneity of TSLP protein preps, a TSLP having the argininewithin the furin cleavage site mutated to alanine was used. See U.S.Pat. No. 7,288,633. Each Bead Code of streptavidin-coated Luminex beads(Luminex, #L100-L1XX-01, XX specifies the bead code) was incubated in100 ul of 6 pg/bead biotinylated monovalent mouse-anti-human IgG captureantibody (BD Pharmingen, #555785) for 1 hour at room temperature in thedark, then washed 3× with PBSA, phosphate buffered saline (PBS) plus 1%bovine serum albumin (BSA). Each bead code was separately incubated with100 ul of a 1:10 dilution anti-TSLP antibody (Coating Antibody) for 1hour then washed. The beads were pooled then dispensed to a 96-wellfilter plate (Millipore, #MSBVN1250). 100 ul of 2 ug/ml parental TSLPwas added to half the wells and buffer to the other half and incubatedfor 1 hour then washed. 100 ul of a 1:10 dilution anti-TSLP antibody(Detection Ab) was added to one well with TSLP and one well withoutTSLP, incubated for 1 hour then washed. An irrelevant human-IgG(Jackson, #009-000-003) as well as a no antibody condition (blank) wererun as negative controls. 20 ul PE-conjugated monovalentmouse-anti-human IgG (BD Pharmingen, #555787) was added to each well andincubated for 1 hour then washed. Beads were resuspended in 75 ul PBSAand at least 100 events/bead code were collected on the BioPlexinstrument (BioRad).

Median Fluorescent Intensity (MFI) of the antibody pair without TSLP wassubtracted from signal of the corresponding reaction containing TSLP.For the antibody pair to be considered bound simultaneously, andtherefore in different bins, the value of the reaction had to meet twocriteria: 1) the values had to be 2 times greater than the coatingantibody paired with itself, the irrelevant or the blank, whichever washighest, and 2) the values had to be greater than the signal of thedetection antibody present with the irrelevant or the blank coated bead.

Analysis of competition between the antibodies was complicated by thefact that there was an incongruity between the performance of antibodiesas probes versus their performance as blockers. However, if oneconsiders only those bins of antibodies that are unambiguous (i.e. eachantibody will block the others when used as a reference) a minimum ofeight bins were found as shown in Table 4 below.

TABLE 4 Bin 1 2 3 4 5 6 7 8 A5 A6 A27 A24 A10 A4 A2 A23 A17 A7 A11 A12A26 A23 A21 A6 A6 A11 A24 A10 A4 A23 A26

It is notable that some antibodies, such as A23 and A6, are found inmultiple bins. It is possible to determine other binning relationships,and the inclusion or exclusion of antibodies from these bins was biasedtowards exclusion.

The results of the assay determined which of the other antibodiescross-compete for binding with the reference antibody. By“cross-competes for binding” it is meant that the reference antibodywhen used as the blocking antibody is able to block binding of the otherantibody when used as a probe and vice versa. In other words, if thereference antibody was able to block the other antibody but the otherantibody was not able to block the reference antibody, the antibodieswere not said to cross-compete. A list of cross-competing antibodies isprovided in Table 5.

TABLE 5 Reference Antibody Exemplary Cross-Competing Antibodies A2  A21,A23 A4  A10, A23, A26 A5  A6, A8, A11, A17 A6  A5, A7, A8, A11, A17, A23A7  A6, A8, A11, A17 A8  A5, A6, A7, A17, A23 A10 A4, A12, A24, A26 A11A5, A6, A7, A17, A24, A27 A12 A10, A24, A26 A17 A5, A6, A7, A8, A11 A21A2, A23, A27 A23 A2, A4, A6, A8, A21 A24 A10, A11, A12, A26, A27 A26 A4,A10, A12, A24 A27 A11, A21, A24

EXAMPLE 9 Epitope Mapping

While epitopes are often thought of as linear sequences, it is moreoften the case that an antibody recognizes a face of the target which iscomposed of discontinuous amino acids. These amino acids may be farapart on the linear sequence but brought close together through thefolding of the target, and antibodies which recognize such an epitopeare known as conformation-sensitive or just conformational antibodies.This kind of binding may be defined through the use of denatured Westernblots, wherein prior to running on a gel the target is heated in thepresence of detergent and reducing agent to unfold it. The blot fromthis gel may then be probed by antibodies, and an antibody which is ableto recognize the target after this treatment probably recognizes alinear epitope. Although the epitopes of antibodies which bind linearsequences may be defined through binding to peptides (e.g. PepSpot),conformational antibodies would not be expected to bind standardpeptides with high affinity.

Reduced, heat-denatured, purified parental TSLP protein was loaded on10% Bis-Tris Nupage gel in MES SDS Running Buffer. Protein wastransferred to PVDF Membrane, blocked with 5% Non-fat Dry Milk (NFDM) inPBS+0.05% Tween (PBST), and incubated with TSLP antibodies for 1 hour atRT. The blots were washed 3× in PBST then incubated with a goatanti-huIgG secondary antibody for 1 hour at RT. The blots were washedagain and incubated with an anti-goat IgG:Alexa 680. After washing 3× inPBST, the blots were scanned on the LiCor to visualize bands.

Antibodies A2, A4, A5, A6, A7, A10, A21, A23, and A26 were characterizedusing this method. Antibodies A2, A4, and A5 bound to the linear epitopeas evidenced by a strong band on the Western Blot. All other antibodieswere conformational as due to no or extremely weak bands on the WesternBlot.

Epitopes may be further defined as structural or functional. Functionalepitopes are generally a subset of the structural epitopes and consistof those residues which directly contribute to the affinity of theinteraction (e.g. hydrogen bonds, ionic interactions). Structuralepitopes may be thought of as the patch of the target which is coveredby the antibody.

Scanning mutagenesis was employed to further define the epitopes boundby the antibodies. Alanine scanning mutagenesis is used frequently todefine functional epitopes; the substitution of alanine (methylsidechain) is essentially an amputation of the wild-type amino acidsidechain and is fairly subtle. Interactions with the protein backbone,such as hydrogen bonding to the amide linkages, would likely not berevealed with alanine scanning. Instead, arginine and glutamic acidscanning mutagenesis was used. These two sidechains were chosen due totheir large steric bulk and their charge, which allows mutations whichoccur in the structural epitope to have a greater effect on antibodybinding. Arginine was generally employed except when the WT reside wasarginine or lysine, and in these cases the residue was mutated toglutamic acid to switch the charge. In a few cases, the WT residue wasmutated to both arginine and glutamic acid.

Ninety-five amino acids, distributed throughout TSLP, were selected formutation to arginine or glutamic acid. As hydrophobic residues aregenerally found inside the folded core of a protein, the selection wasbiased towards charged or polar amino acids to reduce the likelihood ofthe mutation resulting in misfolded protein. As there was no crystalstructure, these residues were chosen essentially at random anddistributed throughout TSLP. As described in Example 8, a TSLPcontaining a mutated furin cleavage site was used.

BIOPLEX™ binding assay was used to measure binding anti-TSLP antibodiesto mutant TSLP. A biotinylated Penta-His Ab (Qiagen, Lot#: 130163339)was bound onto 100 bead codes of streptavidin-coated beads (Luminex,#L100-L1XX-01, XX specifies the bead code). These were used to capturethe his-tagged protein. The 100 bead codes allowed the multiplexing ofall 85 mutants, 3 parental controls, an irrelevant protein and 12blanks. Antibody binding to mutant protein was compared to antibodybinding to the parental.

100 ul of a 1:5 dilution of the TSLP mutants and parental in supernatantand 1 ug/mL purified TSLP WT, 1 ug/mL irrelevant protein or no proteinwere bound to the coated beads for 1 hour at RT with vigorous shaking.The beads were washed and aliquoted into a 96-well filter plate(Millipore). 100 ul anti-TSLP antibodies in 4-fold dilutions were addedto triplicate wells, incubated for 0.5 hours at RT and washed. 100 ul of1:250 dilution of PE-conjugated anti-human IgG Fc (Jackson,#109-116-170) was added to each well, incubated for 0.5 hours andwashed. Beads were resuspended in 75 uL, shaken for at least 3 mins, andread on the BIOPLEX™.

A residue was considered part of the structural epitope (a “hit”) whenmutating it to arginine or glutamic acid disrupted antibody binding.This was seen as a shift in the EC50 or a reduction of maximum signalcompared to antibody binding to parental TSLP.

Statistical analyses of antibody binding curves to parental and mutantswere used to identify statistically significant EC50 shifts. Theanalysis took into consideration variation in the assay and curvefitting.

The EC50s of the mutant binding curves and parental binding curves werecompared. Statistically significant differences were identified as hitsfor further consideration. The curves with “nofit” or “badfit” flagswere excluded from this analysis.

Two sources of variations were considered in the comparison of EC50estimates, variation from the curve fit and the bead-bead variation.Parental and mutants were linked to different beads, hence theirdifference were confounded with the bead-bead difference. The curve fitvariation was estimated by the standard error of the log EC50 estimates.Bead-bead variation was experimentally determined using an experimentwhere parental controls were linked to each one of the beads. The beadvariation in EC50 estimates of parental binding curve were used toestimate the bead-bead variation.

The comparisons of two EC50s (in log scale) were conducted usingStudent's t-test. A t-statistics is calculated as the ratio betweendelta (the absolute differences between EC50 estimates) and the standarddeviation of delta. The variance of delta is estimated by the sum of thethree components, variance estimate of EC50 for mutant and parentalcurves in the nonlinear regression and two times the bead-bead varianceestimated from a separate experiment. The multiple of two for thebead-bead variance is due to the assumption that both mutant andparental beads have the same variance.

The degree of freedom of the standard deviation of delta was calculatedusing the Satterthwaite's (1946) approximation. Individual p-values andconfidence intervals (95% and 99%) were derived based on Student's tdistribution for each comparison. In the case of multiple parentalcontrols, a conservative approach was implemented by picking theparental control that was most similar to the mutant, i.e., picking theones with the largest p-values.

Multiplicity adjustments were important to control the false positiveswhile conducting a large number of tests simultaneously. Two forms ofmultiplicity adjustment were implemented for this analysis: family wiseerror (FWE) control and false discovery rate (FDR) control. The FWEapproach controls the probability that one or more hits are not real;FDR approach controls the expected proportion of false positive amongthe selected hits. The former approach is more conservative and lesspowerful than the latter one. There are many methods available for bothapproaches, for this analysis, Hochberg's (1988) method was chosen forFWE analysis and Benjamini-Hochberg's (1995) FDR method for FDRanalysis. Adjusted p-values for both approaches were calculated eitherfor each antibody or the whole assay.

Mutations whose EC50 was significantly different from parental, i.e.having an FWE adjusted p-value for each antibody of less than 0.01, or amaximal signal below 50% of parental were considered part of thestructural epitope (Table 6). Mutations that were significant by eitherEC50 shirt or max signal reduction for all antibodies were consideredmisfolded. These mutations were; Y15R, T55R, T74R and A77R.

TABLE 6 Summary of mutations that affect antibody binding in the BIOPLEXand are part of the structural epitope. Antibody Linear IncreasedBinding Affinity Decreased Binding Afinity A2  Yes K67E, K97E, K98E,K21E, T25R, S28R, S64R, K73E R100E, K101E, K103E A4  Yes K97E, K98E,R100E, K10E, A14R, K21E, D22R, K73E, K75E, K101E, K103E A76R A5  YesK12E, D22R, S40R, R122E, N124E, R125E, K129E A6  No S40R, S42R, H46R,R122E, K129E A7  No K101E D2R, T4R, D7R, S42R, H46R, T49R, E50R, Q112R,R122E, R125E, K129E A10 No K97E, K98E, R100E, N5R, S17R, T18R, K21E,D22R, T25R, K101E, K103E T33R, H46R, A63R, S64R, A66R, E68R, K73E, K75E,A76R, A92R, T93R, Q94R, A95R A21 No K97E, K98E, R100E, K21E, K21R, D22R,T25R, T33R, S64R, K101E, K103E K73E, K75E, E111R, S114R A23 No K67E,K97E, K98E, E9R, K10E, K12E, A13R, S17R, S20R, R100E, K101E, K103E K21E,K21R, K73E, K75E, N124E, R125E A26 No K97E, K98E, R100E, A14R, K21E,D22R, A63R, S64R, K67E, K101E, K103E K73E, A76R, A92R, A95R

There were several mutations that disrupted the binding of multipleantibodies, notably K73E, K21E, and D22R. The mutagenesis serves toverify the data generated by binning and to further narrow in on theepitope space. The mutations in TSLP appear to affect clusters ofantibodies that bin together.

EXAMPLE 10 Toxicology

Antibodies that bind human TSLP yet also cross-react with TSLP of otherspecies allow for toxicology testing in those species. In this example,an antibody that cross-reacts with cynomolgus monkey TSLP wasadministered to cynomolgus monkeys. The monkeys were then observed fortoxic effects.

A single-dose safety pharmacology study in cynomolgus monkeys indicatedthat a single 300 mg/kg intravenous dose of the antibody had nocardiovascular, respiratory, body temperature, or neurobehavioraleffects.

Cynomolgus monkeys (5/sex/group) were given 30, 100, or 300 mg/kg dosesonce weekly for 4 weeks, subcutaneously. No adverse toxicology wasobserved at any dose. The antibody did not affect clinical observations,body weight, opthalmology, ECGs, clinical pathology or anatomicpathology.

In a separate study, four male telemeterized cynomolgus monkeys weregiven a single intravenous dose of vehicle (day 1) and 300 mg/kgantibody (day 3). Over a four day observation period no effects oncardiovascular, respiratory, or neurological function was observed.

The antibody was further tested to determine cross-reactivity withnormal human and cynomolgus monkey tissue as recommended in the FDAguideline “Points to Consider in the Manufacture and Testing ofMonoclonal Antibody Products for Human Use” (FDA Center for BiologicsEvaluation and Research, 28 Feb. 1997). No staining of normal tissue at1 or 50 ug/mL was observed.

The above results suggest that the antibody is not expected to producetoxic effects in humans.

1. An isolated antigen binding protein comprising an amino acid sequence selected from the group consisting of: a. a light chain CDR3 sequence selected from the group consisting of: i. a light chain CDR3 sequence that differs by no more than a total of two amino acid additions, substitutions, and/or deletions from a CDR3 sequence selected from the group consisting of the light chain CDR3 sequences of A1 to A27; ii. QQAX₈SFPLT; (SEQ ID NO: 251)

and b. a heavy chain CDR3 sequence selected from the group consisting of: i. a heavy chain CDR3 sequence that differs by no more than a total of three amino acid additions, substitutions, and/or deletions from a CDR3 sequence selected from the group consisting of the heavy chain CDR3 sequences of A1 to A27; ii. GGGIX₁₂VADYYX₁₃YGMDV; (SEQ ID NO: 255) and iii. DX₂₁GX₂₂SGWPLFX₂₃Y; (SEQ ID NO: 259)

wherein X₈ is an N residue or a D residue; X₁₂ is a P residue or an A residue; X₁₃ is a Y residue or an F residue; X₂₁ is a G residue or an R residue; X₂₂ is an S residue or a T residue; X₂₃ is an A residue or a D residue. and wherein said antigen binding protein specifically binds to TSLP.
 2. The isolated antigen binding protein of claim 1, further comprising an amino acid sequence selected from the group consisting of: a. a light chain CDR1 sequence selected from the group consisting of: i. a light chain CDR1 sequence that differs by no more than three amino acids additions, substitutions, and/or deletions from a light chain CDR1 sequence of A1-A27; ii. RSSQSLX₁YSDGX₂TYLN; (SEQ ID NO: 246) iii. RASQX₄X₅SSWLA; (SEQ ID NO: 249) and

b. a light chain CDR2 sequence selected from the group consisting of: i. a light chain CDR2 sequence that differs by no more than two amino acid additions, substitutions, and/or deletions from a CDR2 sequence of A1-A27; ii. KVSX₃WDS; (SEQ ID NO: 247) iii. X₆X₇SSLQS; (SEQ ID NO: 250) and iv. QDX₉KRPS; (SEQ ID NO: 252)

c. a heavy chain CDR1 sequence selected from the group consisting of: i. a heavy chain CDR1 sequence that differs by no more than two amino acid additions, substitutions, and/or deletions from a CDR1 sequence of A1-A27; ii. X₁₀YGMH; (SEQ ID NO: 253) and iii. X₁₅X₁₆YMX₁₇; (SEQ ID NO: 257)

d. a heavy chain CDR2 sequence selected from the group consisting of: i. a heavy chain CDR2 sequence that differs by no more than three amino acid additions, substitutions, and/or deletions from a CDR2 sequence of A1-A27; ii. VIWX₁₁DGSNKYYADSVKG; (SEQ ID NO: 254) iii. VISYDGSX₁₄KYYADSVKG; (SEQ ID NO: 256) and iv. WINPNSGGTNX₁₈X₁₉X₂₀KFQG; (SEQ ID NO: 258)

wherein X₁ is a V residue or an I residue; X₂ is an N residue or a D residue; X₃ is a Y residue or an N residue; X₄ is a G residue or a S residue; X₅ is a L residue or an I residue; X₆ is an N residue or a T residue; X₇ is a T residue or an A residue; X₉ is a K residue or an N residue; X₁₀ is an S residue or an N residue; X₁₁ is a Y residue or an F residue; X₁₄ is a Y residue or a N residue; X₁₅ is a D residue or G residue; X₁₆ is a Y residue or a D residue; X₁₇ is a Y residue or an H residue; X₁₈ is a Y residue or an H residue; X₁₉ is a V residue or an A residue; X₂₀ is a Q residue or an R residue, and wherein said antigen binding protein specifically binds to TSLP.
 3. The isolated antigen binding protein of claim 1 comprising either: a. a light chain variable domain comprising: i. a light chain CDR1 sequence selected from A1-A27; ii a light chain CDR2 sequence selected from A1-A27; iii. a light chain CDR3 sequence selected from A1-A27, or b. a heavy chain variable domain comprising: i. a heavy chain CDR1 sequence selected from A1-A27; ii. a heavy chain CDR2 sequence selected from A1-A27, and iii. a heavy chain CDR3 sequence selected from A1-A27; or c. the light chain variable domain of (a) and the heavy chain variable domain of (b).
 4. The isolated antigen binding protein of claim 1 comprising either: a. a light chain variable domain sequence selected from the group consisting of; i. amino acids having a sequence at least 80% identical to a light chain variable domain sequence selected from L1-L27; ii. a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the light chain variable domain sequence of L1-L27; iii. a sequence of amino acids encoded by a polynucleotide sequence that hybridizes under moderately stringent conditions to the complement of a polynucleotide consisting of a light chain variable domain sequence of L1-L27; b. a heavy chain variable domain sequence selected from the group consisting of: i. a sequence of amino acids that is at least 80% identical to a heavy chain variable domain sequence of H1-H27; ii. a sequence of amino acids encoded by a polynucleotide sequence that is at least 80% identical to a polynucleotide sequence encoding the heavy chain variable domain sequence of H1-H27; iii. a sequence of amino acids encoded by a polynucleotide sequence that hybridizes under moderately stringent conditions to the complement of a polynucleotide consisting of a heavy chain variable domain sequence of H1-H27; or c. the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein said antigen binding protein specifically binds to TSLP.
 5. An isolated antigen binding protein, comprising either: a. a light chain variable domain sequence selected from the group consisting of: L1-L27 b. a heavy chain variable domain sequence selected from the group consisting of: H1-H27; or, c. the light chain variable domain of (a) and the heavy chain variable domain of (b), wherein the antigen binding protein specifically binds to TSLP.
 6. The isolated binding protein of claim 5, comprising a light chain variable domain sequence and a heavy chain variable domain sequence selected from the group consisting of: L1H1, L2H2, L3H3, L4H4, L5H5, L6H6, L7H7, L8H8, L9H9, L10H10, L11H11, L12H12, L13.1H13, L13.2H13, L14.1H14, L14.2H14, L15.1H15, L15.2H15, L16.1H16, L16.2H16, L17H17, L18.1H18, L18.2H18, L19.1H19, L19.2H19, L20.1H20, L20.2H20, L21H21, L22H22, L23H23, L24H24, L25H25, L26H26, and L27H27.
 7. The isolated antigen binding protein of claim 1, wherein the binding protein binds to TSLP with substantially the same Kd as a reference antibody selected from the group of antibodies consisting of A2, A3, A4 and A5.
 8. The isolated antigen binding protein of claim 1, wherein the binding protein inhibits TSLP activity according to the primary cell OPG assay with the same IC₅₀ as a reference antibody selected from the group of antibodies consisting of A2, A3, A4, and A5.
 9. The isolated antigen binding protein of claim 1, wherein the antigen binding protein is selected from the group consisting of a human antibody, a humanized antibody, chimeric antibody, a monoclonal antibody, a polyclonal antibody, a recombinant antibody, an antigen-binding antibody fragment, a single chain antibody, a monomeric antibody, a diabody, a triabody, a tetrabody, a Fab fragment, an F(fa′)_(x) fragment, a domain antibody, an IgD antibody, an IgE antibody, and IgM antibody, and IgG1 antibody, and IgG2 antibody, and IgG3 antibody, and IgG4 antibody, and IgG4 antibody having at least one mutation in the hinge region that alleviates a tendency to for intra H-chain disulfide bonds.
 10. The isolated antigen binding protein of claim 1, wherein the antigen binding protein is a human antibody.
 11. A pharmaceutical composition comprising the antibody of claim
 9. 12. An isolated nucleic acid comprising a polynucleotide sequence encoding the light chain variable domain, the heavy chain variable domain, or both, of the antigen binding agent of claim
 5. 13. The isolated nucleic acid of claim 12, wherein the sequence is selected from L1-L27; H1-H27, or both.
 14. A recombinant expression vector comprising the nucleic acid of claim
 12. 15. A host cell comprising the vector of claim
 14. 16. A hybridoma capable of producing the antibody of claim
 10. 17. A method of producing the antibody, comprising incubating the host cell of claim 15 under conditions that allow it to express the antibody.
 18. A method of treating a TSLP-related inflammatory condition in a subject in need of such treatment comprising administering a therapeutically effective amount of composition of claim 11 to the subject.
 19. The method of claim 18, wherein the inflammatory condition is selected from the group consisting of allergic asthma, allergic rhinosinusitis, allergic conjunctivitis, and atopic dermatitis.
 20. A method of treating a TSLP-related fibrotic disorder in a subject in need of such treatment comprising administering a therapeutically effective amount of the composition of claim 11 to the subject.
 21. The method of claim 20, wherein the fibrotic disorder is selected from the group consisting of scleroderma, interstitial lung disease, idiopathic pulmonary fibrosis, fibrosis arising from chronic hepatitis B or C, radiation-induced fibrosis, and fibrosis arising from wound healing.
 22. An isolated antigen binding protein that cross-competes for binding TSLP with an antibody selected from the group consisting of A1-A27.
 23. The isolated antigen binding protein of claim 22, wherein the antigen binding protein comprises an antibody heavy chain variable region and light chain variable region.
 24. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K67E, K97E, K98E, R100E, K101E, and K103E.
 25. The isolated antigen binding protein of claim 24, wherein antigen binding protein has a higher binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 26. The isolated antigen binding protein of claim 25, wherein antigen binding protein has a higher binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 27. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K21E, T25R, S28R, S64R, and K73E.
 28. The isolated antigen binding protein of claim 27, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 29. The isolated antigen binding protein of claim 28, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 30. The isolated antigen binding protein of claim 27, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K67E, K97E, K98E, R100E, K101E, and K103E.
 31. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K97E, K98E, R100E, K101E, and K103E.
 32. The isolated antigen binding protein of claim 31, wherein antigen binding protein has a higher binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 33. The isolated antigen binding protein of claim 32, wherein antigen binding protein has a higher binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 34. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K10E, A14R, K21E, D22R, K73E, K75E, and A76R.
 35. The isolated antigen binding protein of claim 34, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 36. The isolated antigen binding protein of claim 35, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 37. The isolated antigen binding protein of claim 34, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K97E, K98E, R100E, K101E, and K103E.
 38. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K12E, D22R, S40R, R122E, N124E, R125E, and K129E.
 39. The isolated antigen binding protein of claim 38, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 40. The isolated antigen binding protein of claim 39, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 41. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of S40R, S42R, H46R, R122E, and K129E.
 42. The isolated antigen binding protein of claim 41, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 43. The isolated antigen binding protein of claim 42, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 44. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of D2R, T4R, D7R, S42R, H46R, T49R, E50R, Q112R, R122E, R125E, and K129E.
 45. The isolated antigen binding protein of claim 44, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 46. The isolated antigen binding protein of claim 45, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 47. The isolated antigen binding protein of claim 44, wherein the antigen binding protein binds to a mutated TSLP comprising mutation K101E with an affinity higher than the wild-type affinity.
 48. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of N5R, S17R, T18R, K21E, D22R, T25R, T33R, H46R, A63R, S64R, A66R, E68R, K73E, K75E, A76R, A92R, T93R, Q94R, and A95R.
 49. The isolated antigen binding protein of claim 48, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 50. The isolated antigen binding protein of claim 49, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 51. The isolated antigen binding protein of claim 48, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K97E, K98E, R100E, K101E, and K103E.
 52. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K21E, K21R, D22R, T25R, T33R, S64R, K73E, K75E, E111R, and S114R.
 53. The isolated antigen binding protein of claim 52, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 54. The isolated antigen binding protein of claim 53, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 55. The isolated antigen binding protein of claim 52, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K97E, K98E, R100E, K101E, and K103E.
 56. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of E9R, K10E, K12E, A13R, S17R, S20R, K21E, K21R, K73E, K75E, N124E, and R125E.
 57. The isolated antigen binding protein of claim 56, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 58. The isolated antigen binding protein of claim 57, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 59. The isolated antigen binding protein of claim 56, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K67E, K97E, K98E, R100E, K101E, and K103E.
 60. An isolated antigen binding protein that binds wild-type TSLP with a wild-type affinity, wherein the antigen binding protein binds to any of a group of mutated TSLP with an affinity lower than the wild-type affinity, wherein the group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of A 14R, K21E, D22R, A63R, S64R, K67E, K73E, A76R, A92R, and A95R.
 61. The isolated antigen binding protein of claim 60, wherein antigen binding protein has a lower binding affinity for any two or more members of the group of mutated TSLP than the wild-type affinity.
 62. The isolated antigen binding protein of claim 61, wherein antigen binding protein has a lower binding affinity for all members of the group of mutated TSLP than the wild-type affinity.
 63. The isolated antigen binding protein of claim 60, wherein the antigen binding protein binds to any of a second group of mutated TSLP with an affinity higher than the wild-type affinity, wherein the second group of mutated TSLP includes mutated TSLP comprising a mutation selected from the group consisting of K97E, K98E, R100E, K101E, and K103E. 